WO2010080223A1

WO2010080223A1 - System and method for preparing samples

Info

Publication number: WO2010080223A1
Application number: PCT/US2009/065338
Authority: WO
Inventors: David J. Velasquez; Cynthia D. Zook; Jon A. Kirschhoffer
Original assignee: 3M Innovative Properties Company
Priority date: 2008-12-18
Filing date: 2009-11-20
Publication date: 2010-07-15

Abstract

A system and method for preparing samples. The system (100) can include a first container (102) adapted to contain a liquid composition comprising a source, and a second container (104) adapted to be coupled to the first container. The first container (102) can be collapsible and can be adapted to allow the liquid composition to be agitated. The second container (104) can be adapted to receive at least a portion of the liquid composition from the first container (102), and can be further adapted to enrich a microorganism of interest in the liquid composition to form an enriched liquid composition. The method can include providing a first container containing a source, agitating the source to form a liquid composition, and moving at least a portion of the liquid composition from the first container to a second container coupled to the first container.

Description

SYSTEM AND METHOD FOR PREPARING SAMPLES

FIELD The present disclosure generally relates to a system and method for preparing samples, and particularly, for preparing samples and testing the samples for one or more microorganisms .

BACKGROUND

In a variety of applications, food and non-food sources may need to be tested for microorganisms (e.g., bacteria, viruses, fungi, spores, etc.) and/or other analytes of interest

(e.g., toxins, allergens, hormones, etc.). For example, foods grown, purchased and consumed by the general population may contain or acquire microorganisms or other analytes, which can flourish or grow as a function of the environment in which they are located. This growth may lead to accelerated spoilage of the food product or to the proliferation of pathogenic organisms, which may produce toxins or multiply to infective doses. By way of further example, a variety of analytical methods can be performed on samples of non-food sources (e.g., groundwater, urine, etc.) to determine if the sample contains a particular analyte. For example, groundwater can be tested for a microorganism or a chemical toxin; and urine can be tested for a variety of diagnostic indicators to enable a diagnosis (e.g., diabetes, pregnancy, etc.).

SUMMARY

One aspect of the present disclosure provides a sample preparation system. The system can include a first container adapted to contain a liquid composition comprising a source, and a second container adapted to be coupled to the first container. The first container can be collapsible and can be adapted to allow the liquid composition to be agitated. The second container can be adapted to receive at least a portion of the liquid composition from the first container, and can further be adapted to enrich a microorganism of interest in the liquid composition to form an enriched liquid composition.

Another aspect of the present disclosure provides a sample preparation system. The system can include a first container containing a liquid composition comprising a source, and a second container coupled to the first container. The first container can be collapsible. The second container can contain at least a portion of the liquid composition, and the second container can be adapted to enrich a microorganism of interest in the liquid composition to form an enriched liquid composition.

Another aspect of the present disclosure can provide a method for preparing samples. The method can include providing a first container containing a source, and agitating the source in the first container to form a liquid composition comprising the source. The first container can be collapsible. The method can further include moving at least a portion of the liquid composition from the first container to a second container coupled to the first container, the second container adapted to enrich a microorganism of interest in the liquid composition to form an enriched liquid composition.

Other features and aspects of the present disclosure will become apparent by consideration of the detailed description and accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic flow chart depicting a sample preparation method according to one embodiment of the present invention.

FIG. 2 is a schematic flow chart depicting a sample preparation method according to another embodiment of the present invention.

FIG. 3 is a schematic plan view of a sample preparation system according to one embodiment of the present invention.

FIG. 4 is a schematic plan view of a sample preparation system according to another embodiment of the present invention.

FIG. 5 is a schematic perspective view of a sample preparation system according to another embodiment of the present disclosure.

FIG. 6 is a schematic side elevational view of the sample preparation system of

FIG. 5. FIG. 7 is a schematic perspective view of a sample preparation system according to another embodiment of the present disclosure.

FIG. 8 is a schematic side elevational view of the sample preparation system of FIG. 7.

FIG. 9 is a schematic front perspective view of the sample preparation system according to another embodiment of the present disclosure, the sample preparation system shown in a first configuration.

FIG. 10 is a schematic rear perspective view of the sample preparation system of FIG. 9 in a second configuration.

FIG. 11 is a schematic perspective view of a sample preparation system according to another embodiment of the present disclosure.

DETAILED DESCRIPTION

Before any embodiments of the present disclosure are explained in detail, it is to be understood that the invention is not limited in its application to the details of construction and the arrangement of components set forth in the following description or illustrated in the following drawings. The invention is capable of other embodiments and of being practiced or of being carried out in various ways. Also, it is to be understood that the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting. The use of "including," "comprising," or "having" and variations thereof herein is meant to encompass the items listed thereafter and equivalents thereof as well as additional items. Unless specified or limited otherwise, the term "coupled," "connected," and variations thereof are used broadly and encompass both direct and indirect couplings and connections. Further, "coupled" is not restricted to physical or mechanical couplings. It is to be understood that other embodiments may be utilized, and structural or logical changes may be made without departing from the scope of the present disclosure. Furthermore, terms such as "front," "rear," "upper," "lower," "left," "right," and the like are only used to describe elements as they relate to one another, but are in no way meant to recite specific orientations of the apparatus, to indicate or imply necessary or required orientations of the apparatus, or to specify how the invention described herein will be used, mounted, displayed, or positioned in use.

The present disclosure generally relates to a sample preparation system, for example, a sample preparation system used to prepare and test samples for the presence, quantity, and/or viability of one or more microorganisms. The present disclosure also generally relates to a method for using a sample preparation system to prepare a sample and test the sample for the presence, quantity, and/or viability of one or more microorganisms .

The sample preparation system and method can be used to prepare a sample from a liquid composition for subsequent processing (e.g., enrichment) and/or testing. Such a liquid composition can be obtained in a variety of ways. For example, in some embodiments, the source of interest is itself a liquid and makes up the liquid composition. In some embodiments, the liquid composition can include the liquid resulting from washing or rinsing a source of interest (e.g., a surface, fomite, etc.) with a diluent. In some embodiments, the liquid composition can include the resulting liquid mixture after combining a source and a diluent (either before or after the source and the diluent are added to the sample preparation system). In some embodiments, the liquid composition can include the filtrate resulting from filtering a liquid composition resulting from combining a source of interest with an appropriate diluent. That is, in some embodiments, large insoluble matter, such as various foods, fomites, or the like, can be removed from a liquid composition in a first filtration step. The filtrate resulting from such a first filtration step can then be further mixed, enriched, etc. with the sample preparation system and method of the present disclosure. In addition, such a first filtration step can be adapted to allow the analyte(s) of interest, if present, to remain in the filtrate, such that the analyte(s) of interest will pass on to the subsequent steps in the sample preparation method. Such a first filtration step can occur prior to introducing the liquid composition to the sample preparation system, or it can occur after the liquid composition has been added to the sample preparation system (e.g., such a first filtration step can occur within the sample preparation system). The term "source" is generally used to refer to the food or nonfood desired to be tested for analytes. The source can be a solid, a liquid, a semi-solid, a gelatinous material, and combinations thereof. In some embodiments, the source can be provided by a substrate that was used, for example, to collect the source from a surface of interest. In some embodiments, the source can include the substrate, which can be further broken apart (e.g., during an agitation or dissolution process) to enhance retrieval of the source and any microorganism of interest from the substrate. The surface of interest can include at least a portion of a variety of surfaces, including, but not limited to, walls (including doors), floors, ceilings, drains, refrigeration systems, ducts (e.g., airducts), vents, toilet seats, handles, doorknobs, handrails, bedrails (e.g., in a hospital), countertops, tabletops, eating surfaces (e.g., trays, dishes, etc.), working surfaces, equipment surfaces, clothing, etc., and combinations thereof. All or a portion of the source can be used in the sample preparation system and method. When a portion of the source is used, this can sometimes be referred to as a "sample" of the source. However, the term "sample" is generally used herein to refer to a volume or mass of material that is extracted from, or moved within, the sample preparation system for further analysis (e.g., detection of analytes). A "sample" can include all or a portion of a source.

The term "food" is generally used to refer to a solid, liquid (e.g., including, but not limited to, a solution, a dispersion, a emulsion, a suspension, etc., and combinations thereof) and/or semi-solid comestible composition. Examples of foods include, but are not limited to, meats, poultry, eggs, fish, seafood, vegetables, fruits, prepared foods (e.g., soups, sauces, pastes), grain products (e.g., flour, cereals, breads), canned foods, milk, other dairy products (e.g., cheese, yogurt, sour cream), fats, oils, desserts, condiments, spices, pastas, beverages, water, animal feed, other suitable comestible materials, and combinations thereof.

The term "nonfood" is generally used to refer to sources of interest that do not fall within the definition of "food" and are generally not considered to be comestible. Examples of nonfood sources can include, but are not limited to, clinical samples, cell lysates, whole blood or a portion thereof (e.g., serum), other bodily fluids or secretions (e.g., saliva, sweat, sebum, urine), feces, cells, tissues, organs, biopsies, plant materials, wood, soil, sediment, medicines, cosmetics, dietary supplements (e.g., ginseng capsules), pharmaceuticals, fomites, other suitable non-comestible materials, and combinations thereof.

The term "fomite" is generally used to refer to an inanimate object or substrate capable of carrying infectious organisms and/or transferring them. Fomites can include, but are not limited to, cloths, mop heads, towels, sponges, wipes, eating utensils, coins, paper money, cell phones, clothing (including shoes), doorknobs, feminine products, diapers, etc., portions thereof, and combinations thereof.

A source can be tested for the presence or absence of particular microorganisms, one or more microorganisms can be quantitated, and/or the source can be tested for the viability of one or more microorganisms. Such microorganisms can be present within a source (e.g., on the interior), or on the exterior (e.g., on the outer surface) of a source. In some embodiments, testing for the presence or absence of a microorganism of interest can include testing for an analyte that is indicative of the presence, quantity, and/or viability of a microorganism. The term "analyte" is generally used to refer to a substance to be detected (e.g., by a laboratory or field test). Examples of such analytes can include, but are not limited to, biomolecules, chemicals, metal ions (e.g. mercury ions, heavy metal ions), metal-ion-containing complexes (e.g., complexes comprising metal ions and organic ligands), and combinations thereof.

The term "microorganism" is generally used to refer to any prokaryotic or eukaryotic microscopic organism, including without limitation, one or more of bacteria (e.g., motile or vegetative, Gram positive or Gram negative), viruses (e.g., Norovirus, Norwalk virus, Rotavirus, Adenovirus, DNA viruses, RNA viruses, enveloped, non- enveloped, human immunodeficiency virus (HIV), human Papillomavirus (HPV), etc.), bacterial spores or endospores, algae, fungi (e.g., yeast, filamentous fungi, fungal spores), prions, mycoplasmas, parasites, and protozoa. In some cases, the microorganisms of particular interest are those that are pathogenic, and the term "pathogen" is used to refer to any pathogenic microorganism. Examples of pathogens can include, but are not limited to, members of the family Enterobacteriaceae, or members of the family Micrococaceae, or the genera Staphylococcus spp., Streptococcus, spp., Pseudomonas spp., Enterococcus spp., Salmonella spp., Legionella spp., Shigella spp., Yersinia spp., Enterobacter spp., Escherichia spp., Bacillus spp., Listeria spp., Campylobacter spp., Acinetobacter spp.,

Vibrio spp., Clostridium spp., and Corynebacterium spp. Particular examples of pathogens can include, but are not limited to, Escherichia coli including enterohemorrhagic E. coli e.g., serotype O157:H7, O129:H11; Pseudomonas aeruginosa; Bacillus cereus; Bacillus anthracis; Salmonella enteritidis; Salmonella enterica serotype

Typhimurium; Listeria monocytogenes; Clostridium botulinum; Clostridium perfringens;

Staphylococcus aureus; methicillin-resistant Staphylococcus aureus; Campylobacter jejuni; Yersinia enterocolitica; Vibrio vulnificus; Clostridium difficile; vancomycin- resistant Enterococcus; and Enterobacter [Cronobacter] sakazakii. Environmental factors that may affect the growth of a microorganism (e.g., that can be used in a sample preparation system of the present disclosure) can include the presence or absence of nutrients, pH, moisture content, oxidation-reduction potential, antimicrobial compounds, temperature, atmospheric gas composition and biological structures or barriers.

A variety of testing methods can be used to identify, quantitate, and/or elucidate the viability of one or more microorganisms, including, but not limited to, microbiological assays, biochemical assays (e.g. immunoassay), or a combination thereof. Specific examples of testing methods that can be used include, but are not limited to, lateral flow assays, titration, thermal analysis, microscopy (e.g., light microscopy, fluorescent microscopy, immunofluorescent microscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM)), spectroscopy (e.g., mass spectroscopy, nuclear magnetic resonance (NMR) spectroscopy, Raman spectroscopy, infrared (IR) spectroscopy, x-ray spectroscopy, attenuated total reflectance spectroscopy, Fourier transform spectroscopy, gamma-ray spectroscopy, etc.), spectrophotometry (e.g., absorbance, fluorescence, luminescence, etc.), chromatography (e.g., gas chromatography, liquid chromatography, ion-exchange chromatography, affinity chromatography, etc.), electrochemical analysis, genetic techniques (e.g., polymerase chain reaction (PCR), transcription mediated amplification (TMA), hybridization protection assay (HPA), DNA or RNA molecular recognition assays, etc.), adenosine triphosphate (ATP) detection assays, immunological assays (e.g., enzyme-linked immunosorbent assay (ELISA)), cytotoxicity assays, viral plaque assays, techniques for evaluating cytopathic effect, culture techniques such as those that can be done using a growth medium (e.g., agar) and/or 3M™ PETRIFILM™ plates (e.g., and imaged, quantitated and/or interpreted using a 3M™ PETRIFILM™ plate reader (3M Company, St. Paul, MN)), other suitable analyte testing methods, or a combination thereof.

The term "biomolecule" is generally used to refer to a molecule, or a derivative thereof, that occurs in or is formed by an organism. For example, a biomolecule can include, but is not limited to, at least one of an amino acid, a nucleic acid, a polypeptide, a protein, a polynucleotide, a lipid, a phospholipid, a saccharide, a polysaccharide, and combinations thereof. Specific examples of biomolecules can include, but are not limited to, a metabolite (e.g., staphylococcal enterotoxin), an allergen (e.g., peanut allergen(s), egg allergen(s), pollens, dust mites, molds, danders, or proteins inherent therein, etc.), a hormone, a toxin (e.g., Bacillus diarrheal toxin, aflatoxin, Clostridium difficile toxin etc.),

RNA (e.g., mRNA, total RNA, tRNA, etc.), DNA (e.g., plasmid DNA, plant DNA, etc.), a tagged protein, an antibody, an antigen, ATP, and combinations thereof.

The terms "soluble matter" and "insoluble matter" are generally used to refer to matter that is relatively soluble or insoluble in a given medium, under certain conditions. Specifically, under a given set of conditions, "soluble matter" is matter that goes into solution and can be dissolved in the solvent (e.g., a diluent) of a system. "Insoluble matter" is matter that, under a given set of conditions, does not go into solution and is not dissolved in the solvent of a system. A source can include soluble matter and insoluble matter (e.g., cell debris). Insoluble matter is sometimes referred to as particulate(s) or debris and can include portions of the source material itself (i.e., from internal portions or external portions (e.g., the outer surface) of the source) or other source residue or debris resulting from an agitation process. The microorganism of interest can be present in the soluble matter or the insoluble matter.

The term "filtering" is generally used to describe the process of separating matter by size, density, charge and/or function. For example, filtering can include separating soluble matter and a solvent (e.g., diluent) from insoluble matter, or it can include separating soluble matter, a solvent and relatively small insoluble matter from relatively large insoluble matter, etc. A variety of filtration methods can be used, including, but not limited to, passing the source through a filter, settling followed by aspiration or decanting, other suitable filtration methods, and combinations thereof. "Settling" is used to refer to allowing the more dense matter in the liquid composition (i.e., the matter having a higher density than the diluent and other matter in the mixture) to settle. Settling may occur by gravity or by centrifugation. The more dense matter can then be separated from the less dense matter (and diluent) by aspirating the less dense (i.e., unsettled or floating) and diluent from the more dense matter, decanting the less dense matter and diluent, or a combination thereof. Pre-settling steps can be used in addition to or in lieu of pre-filtering steps to obtain a sample that is to be processed using the sample processing systems and methods of the present disclosure.

A "filter" is generally used to describe the device used to separate the soluble matter (or soluble matter and relatively small insoluble matter) and solvent from the insoluble matter (or relatively large insoluble matter) in a liquid composition. Examples of filters can include, but are not limited to, a woven or non-woven mesh (e.g., a wire mesh, a cloth mesh, a plastic mesh, etc.), a woven or non-woven polymeric web (e.g., comprising polymeric fibers laid down in a uniform or nonuniform process, which may or may not be calendered), a sieve, glass wool, a frit, filter paper, foam, etc., and combinations thereof.

The term "filtrate" is generally used to describe the liquid remaining after the insoluble matter (or at least the relatively large insoluble matter) has been removed from a liquid composition in a filtering process. Because filtering includes a broad range of methods, the term "filtrate" can also be used to refer to the supernatant that results from allowing more dense matter in a mixture to settle. Such a filtrate can include the analyte(s) of interest, if present, and can be passed to subsequent processing, sampling and/or testing steps. Alternatively, the term "filtrate" can be used to describe the undesirable portions of a liquid composition that are removed in an effort to increase the concentration of the analyte(s) of interest in the liquid composition.

The term "enrich" is generally used to refer to the process of growing one or more microorganisms of interest and/or suppressing (e.g., suppressing the growth of and/or killing) one or more non-target microorganisms. "Growing" can include growing in number, and generally refers to increasing the cell population of the microorganism of interest.

The term "rigid" is generally used to refer to a material that is generally not compressible or deformable (e.g., by hand, by a paddle blender, etc.), and particularly, to a material that is generally not compressible or deformable (e.g., by hand, by a paddle blender, etc.) without rupture.

The term "collapsible" is generally used to refer to an object that collapses or deforms under its own weight. For example, a bag may be collapsible. The term "deformable" is generally used to refer to a structure that can be altered from its original shape or state by pressure (e.g., positive or negative) or stress. In some embodiments, the sample preparation system can include a deformable container to which pressure can be applied to reduce its size from its original (i.e., unstressed) dimensions. Such pressure can be used to promote movement of a source from one container to another.

The term "freestanding" is generally used to refer to an object that is capable of standing on its own without being held by another object. The term "self-supporting" is generally used to refer to an object that does not collapse or deform under its own weight.

For example, a bag may not be "self-supporting" in that it may not maintain its shape under its own weight, but rather collapses or distorts. A self-supporting material or object is not necessarily rigid, and may be deformable (e.g., may be compressed or deformed by hand, by a paddle blender, etc.). A self-supporting object is not necessarily freestanding, and a freestanding object is not necessarily self-supporting.

The term "agitate" and derivatives thereof is generally used to describe the process of giving motion to a source (and, in some embodiments, a liquid composition comprising the source and, optionally, diluent(s)), for example, to break up, homogenize, mix, combine and/or blend the contents of the source or such liquid composition, or to liquefy a solid source by blending with a diluent. A variety of agitation methods can be used, including, but not limited to, manual shaking, mechanical shaking (e.g., linear shaking), sonic (e.g., ultrasonic) vibration, vortex stirring, mechanical stirring (e.g., by a magnetic stirbar, or another agitating aid, such as ball bearings), manual compression (e.g., manual beating, squeezing, kneading, pummeling, etc., and combinations thereof), mechanical compression (e.g., paddle blending, mechanical beating, squeezing, kneading, pummeling, etc., and combinations thereof), and combinations thereof.

A variety of agitating devices and processes can be used to break up and/or homogenize a source of material. One example of a suitable agitating process is paddle blending. The phrase "paddle blending" is generally used to refer to a process of pummeling a source (e.g., within a bag) with one or more paddles, in order to break up and/or homogenize the source. Examples of paddle blenders include, but are not limited to, STOMACHER® lab blenders (available from Seward Laboratory System, Inc., Bohemia, NY; and generally described in US Patent No. 3,819,158 (Sharpe et al), which is incorporated herein by reference), which are paddle blending devices that include two or more paddles which alternately push on a bag containing a source, causing the source to move around and break up within the bag. Some existing bags that can be used with a paddle blender can be similar to plastic storage bags with one end open to receive a source. Some existing bags are sized and configured to be placed into a compartment in a paddle blender, and some existing paddle blenders include a mechanism that is adapted to close and seal the bag within the compartment of the paddle blender, and are also adapted to maintain the bag in the proximity of the paddles of the paddle blender during the paddle blending process.

In some existing paddle blending systems, after agitating a source in a bag, a sample of the agitated source (or liquid composition) can be removed (e.g., by pipetting), and transferred to another container for further processing, or to an assay system. Further processing can include one or more of dilution, enrichment in appropriate media, and the like. After a sample has been transferred, the bag and its contents can be discarded. Some existing bags adapted for use in a paddle blender incorporate a mesh filter pocket. A source of interest (e.g., food) can be positioned inside the mesh filter pocket, or outside the mesh filter pocket. After paddle blending, a pipette can be inserted into the bag (e.g., inside or outside the mesh, i.e., on the opposite side of the mesh from the source) to remove a portion of the filtrate. The sample transfer steps in some of the existing paddle blending devices described above can lead to contamination of the sample and/or the environment because of the collapsible nature of the bag, the openness of the bag, its lack of supporting structure, its inability to be freestanding, and/or the potential difficulty in accessing the source at the bottom of the bag through the open top of the bag. That is, in many existing systems, the bag size is constant to ensure proper closure of the bag by the paddle blender and proper positioning of the bag near the paddles. When small-volume sources are used, however, accessing the source at the bottom of the bag for further processing can be cumbersome.

In contrast, the sample preparation systems of the present disclosure include one or more additional (or secondary) containers adapted to be coupled to a primary container and adapted to enrich at least one microorganism of interest. The secondary container can be further adapted to facilitate transfer of at least a portion of the source (or a liquid composition comprising the source) from the primary container to the one or more secondary containers to minimize contamination and/or to increase transfer efficiency.

The sample preparation system of the present disclosure allows for samples not only to be prepared in a convenient manner but, in some embodiments, also allows for further processing (e.g., enrichment) within a single disposable construction which can maintain integrity of the source and can greatly reduce the possibility of contamination. At least a portion of a source, a liquid composition comprising the source, an enriched liquid composition, and/or a filtrate of any of the above, can be transferred from one portion of the sample preparation system to another, or to an external receiver system, such as an input to an assay system (also sometimes referred to as a "detection device").

For example, in some embodiments, the sample preparation system can be designed to couple (e.g., directly) to a variety of assay systems, including, but not limited to, agar plates or 3M™ PETRIFILM™ plates (available from 3M Company, St. Paul,

MN), lateral flow immunoassays, molecular genetic detection systems, other suitable assay systems capable of performing any of the above-described testing methods, or combinations thereof. By configuring the sample preparation system of the present disclosure to be able to be coupled to an assay system, the need for a manual transfer step to downstream systems can be eliminated, which can further reduce the opportunity for sample or environmental contamination.

FIG. 1 illustrates a schematic flow chart of a sample preparation method 10 according to one embodiment of the present disclosure. The sample preparation method 10 can begin with obtaining a source 12. A diluent 13 can be combined with all or a portion of the source 12 and agitated to form a liquid composition 15 comprising the source 12 dissolved, dispersed, suspended, or emulsified in the diluent 13. As such, the liquid composition 15 is generally a mixture, and can be a solution, an emulsion, a dispersion, a suspension, or a combination thereof. A sample 20 can be removed from the liquid composition 15 for analysis and/or further processing. The liquid composition 15 can include the source 12, the source 12 and any diluent(s) present in the source 12 itself, or the source 12 and one or more diluents 13 that are added to the source 12.

The liquid composition 15 can include soluble matter and insoluble matter, such that some portions (e.g., the microorganism(s) and/or analyte(s) of interest) of the source 12 may be dissolved in the diluent 13 (if employed), while other portions of the source 12 are suspended, dispersed or emulsified in the diluent 13. The liquid composition 15 can be filtered to form a filtrate, and the sample 20 (or the portion moved and enriched) can, in some embodiments, include all or a portion of the liquid composition or the filtrate (e.g., in embodiments in which the liquid composition 15 is filtered). For simplicity, the phrase "liquid composition" will be used to refer to a liquid composition as described above, or a filtrate thereof.

The diluent 13 is generally a liquid and, in some embodiments, is a sterile liquid. In some embodiments, the diluent 13 can include a variety of additives, including, but not limited to, surfactants, or other suitable additives that aid in dispersing, dissolving, suspending or emulsifying the source for subsequent analyte testing; rheological agents; antimicrobial neutralizers (e.g., that neutralize preservatives or other antimicrobial agents); enrichment or growth medium comprising nutrients (e.g., that promote selective growth of desired microorganism(s)) and/or growth inhibitors (e.g., that inhibit the growth of undesired microorganism(s)); pH buffering agents; enzymes; indicator molecules (e.g. pH or oxidation/reduction indicators); spore germinants; an agent to neutralize sanitizers (e.g., sodium thiosulfate neutralization of chlorine); an agent intended to promote bacterial resuscitation (e.g., sodium pyruvate); stabilizing agents (e.g., that stabilize the analyte(s) of interest, including solutes, such as sodium chloride, sucrose, etc.); or a combination thereof. In some embodiments, the diluent 13 includes sterile water (e.g., sterile double- distilled water (ddH₂O)); one or more organic solvents to selectively dissolve, disperse, suspend, or emulsify the source; aqueous organic solvents, or a combination thereof. In some embodiments, the diluent 13 is a sterile buffered solution (e.g., Butterfield's Buffer, available from Edge Biological, Memphis TN). In some embodiments, the diluent 13 is a selective or semi-selective nutrient formulation, such that the diluent 13 may be used in the selective or semi-selective growth of the desired analyte(s) (e.g., bacteria). In such embodiments, the diluent 13 can be incubated with the source 12 for a period of time (e.g., at a specific temperature) to promote such growth of the desired analyte(s).

Examples of growth medium can include, but are not limited to, Tryptic Soy Broth (TSB), Buffered Peptone Water (BPW), Universal Pre-enrichment Broth (UPB), Listeria Enrichment Broth (LEB), Lactose Broth, Bolton broth, or other general, non-selective, or mildly selective media known to those of ordinary skill in the art. The growth medium can include nutrients that support the growth of more than one desired microorganism (i.e., analyte of interest).

Examples of growth inhibitors can include, but are not limited to, bile salts, sodium deoxycholate, sodium selenite, sodium thiosulfate, sodium nitrate, lithium chloride, potassium tellurite, sodium tetrathionate, sodium sulphacetamide, mandelic acid, selenite cysteine tetrathionate, sulphamethazine, brilliant green, malachite green oxalate, crystal violet, Tergitol 4, sulphadiazine, amikacin, aztreonam, naladixic acid, acriflavine, polymyxin B, novobiocin, alafosfalin, organic and mineral acids, bacteriophages, dichloran rose bengal, chloramphenicol, chlortetracycline, certain concentrations of sodium chloride, sucrose and other solutes, and combinations thereof.

In some embodiments, the source 12 includes the diluent 13. For example, a food source that includes a substantial amount of water or other liquid can be agitated to form a liquid composition without adding additional diluent. In some embodiments, the source 12 may be completely dissolved in the diluent 13, such that the liquid composition 15 includes a minimal amount of insoluble matter 15, making any filtering of the liquid composition 15 unnecessary.

In some embodiments, at least a portion of the liquid composition 15 is transferred to another container and enriched to make a first enriched liquid composition 25. The first enriched liquid composition 25 can be enriched for one or more microorganisms of interest. A sample 30 of the first enriched liquid composition 25 (i.e., all or a portion of the first enriched liquid composition 25) can be removed for analysis and/or further processing.

As shown in FIG. 1, in some embodiments, all or a portion of the first enriched liquid composition 25 can be transferred and further enriched for the same or a different microorganism of interest to create a second enriched liquid composition 35. A sample 40 of the second enriched liquid composition 35 (i.e., all or a portion of the second enriched liquid composition 35) can be removed for analysis and/or further processing. Similar to the liquid composition, the first and second enriched liquid compositions 25 and 35 can be filtered to form a filtrate, and the samples 30 and 40, respectively, (or the portion(s) moved and enriched) can, in some embodiments, include all or a portion of the enriched liquid composition 25 or 35, or the respective filtrate. For simplicity, the phrase "enriched liquid composition" will be used to refer to an enriched liquid composition or a filtrate thereof.

FIG. 2 illustrates a schematic flow chart of a sample preparation method 10' according to another embodiment of the present disclosure, wherein like numerals refer to like elements. As shown in FIG. 2, in some embodiments, at least a portion of the liquid composition 15' can be transferred to two secondary containers to form a first enriched liquid composition 25' and a second enriched liquid composition 35' in parallel. Such parallel processing can be used, for example, when the source 12' is to be tested for more than one microorganism, and the source 12' or liquid composition 15' needs to be enriched for more than one microorganism, for example, with different enrichment media or under different enrichment conditions. In some embodiments, the sample preparation method of the present disclosure can include a combination of the sample preparation method 10 shown in FIG. 1, wherein a first enriched liquid composition 25 is formed in series with a second enriched liquid composition 35, and the sample preparation method 10' shown in FIG. 2, where a first enriched liquid composition 25' is formed in parallel with a second enriched liquid composition 35'.

Each of the sample preparation method 10 and 10' are illustrated as including a first enriched liquid composition 25, 25' and a second enriched liquid composition 35, 35'; however, it should be understood that the sample preparation methods 10 and 10' are shown by way of example only, and the sample preparation method 10, 10' need not include a second enriched liquid composition 35, 35', and that the second enriched liquid composition 35, 35' is illustrated in FIGS. 1 and 2 merely to illustrate the serial and parallel processing that can be employed when more than one secondary enrichment step (and secondary container) is employed.

FIG. 3 illustrates a schematic representation of a sample preparation system 100 according to one embodiment of the present disclosure. As shown in FIG. 3, the sample preparation system 100 includes a first container 102 adapted to receive and contain a source, a second container 104 adapted to be coupled to the first container 102, and further adapted to enrich a first microorganism of interest in a liquid composition comprising the source, and a third container 106. The third container 106 can also be adapted to be coupled to at least one of the first container 102 and the second container 104, and can also be further adapted to enrich a second microorganism of interest in a liquid composition comprising the source. The second microorganism of interest can be the same as or different from the first microorganism of interest. The first container 102 can be considered a "primary container," and both the second container 104 and the third container 106 can be considered "secondary containers" in which enrichment can occur.

In some embodiments, as shown in FIG. 3, the containers 102, 104 and 106 are separate and distinct containers. In such embodiments, the source can be positioned in one container 102, 104, 106, agitated to form a liquid composition (and, optionally, a diluent can be added), and then all or a portion of the liquid composition can be moved to another container 102, 104, 106. For example, when all or a portion of the liquid composition is moved from the first container 102 to the second container 104, that portion of the liquid composition resides in the second container 104 and no longer resides in the first container 102.

In some embodiments, one or more of the containers 102, 104, 106 can be adapted to withstand compression. For example, in some embodiments, the first container 102 and the second container 104 are desired to be separate and distinct, and used for separate processing steps. In such embodiments, it can be desirable for the first and second containers 102, 104 not to intermix or share contents (e.g., after the desired material transfer has taken place). As such, it can be desirable for both the first container 102 and the second container 104 to be adapted to withstand compression, for example, by being formed of appropriate materials, by configuring any seals thereon to have sufficient strength, etc. Such compression can include any of the above described manual and mechanical compression techniques.

As shown in FIG. 3, the sample preparation system 100 further includes a first port

(or aperture) 122 positioned to provide fluid communication between ambience and the interior of the first container 102. In addition, the sample preparation system 100 includes a second port 124 and a third port 126 positioned to provide fluid communication between ambience and the interior of the second container 104 and the third container 106, respectively.

In some embodiments, each of the ports 122, 124, and 126 can include a barrier 123, 127, 129 (e.g., a frangible barrier, a removable barrier (e.g., a film, a cap, etc.), or combinations thereof), respectively, positioned over or in the port 122, 124, 126 to inhibit the respective container 102, 104, 106 from being opened to ambience prematurely. Such a barrier 123, 127, 129 can be adapted to be opened by a variety of means, such as by the puncturing force of a pipette tip, a syringe tip, or the like, by removing the barrier, or a combination thereof. One of ordinary skill in the art will understand, however, that a variety of other mechanisms can be employed to seal the port 122, 124, 126 (e.g., a liquid- tight seal, a hermetic seal, or a combination thereof), such that the sample preparation system 100 is inhibited from leaking under normal operating conditions. In some embodiments, each of the ports 122, 124 and 126 can include a valve that can be opened by another device, either temporarily or permanently, such as by a pipette tip, a syringe tip, or the like. The ports 122, 124 and 126 are adapted to allow for physical transfer (e.g., manual and/or automatic) of a liquid composition from the first container 102 to one or both of the second container 104 and the third container 106. In some embodiments, at least a portion of the liquid composition can be moved from the first container 102 into a transfer device (e.g., a pipette, syringe, or the like) and then moved from the transfer device into one or both of the second container 104 and the third container 106. Alternatively, or in addition, a connector (e.g., a tube that is positioned at least partially externally with respect to the first container 102 and/or the second and third containers 104 and 106) can be coupled from the first port 122 to the second port 124 and/or the third port 126 to allow at least a portion of the source to be transferred between containers 102, 104, 106.

Each port 122, 124, 126 can be positioned to allow access to the interior of the respective container 102, 104, 106 for liquid composition retrieval, while minimizing the risk of environmental contamination or of losing or contaminating the liquid composition.

In some embodiments, one or both of the second container 104 and the third container 106 can be integrally formed with the first container 102. In addition, in some embodiments, one or both of the second container 104 and the third container 106 can be adapted to be coupled to the first container 102 and can be removable from the first container 102, such that the second container 104 and/or the third container 106 can be removed from the first container 102 for further processing (e.g., incubation) and/or assaying, for example, after at least a portion of the liquid composition has been transferred to the second container 104 and/or third container 106. In embodiments in which the second container 104 and the third container 106 are removable from the first container 102, the second container 104 and/or the third container 106 can be coupled to the first container 102 at any point in the process. For example, in some embodiments, one or both of the second container 104 and the third container 106 can be coupled to the first container 102 before or after the source is added to the first container 102, such that one or both of the second and third containers 104 and 106 are coupled to the first container 102 during the time that the source is agitated in the first container 102. In some embodiments, the second and third containers 104 and 106 can be coupled to the first container 102 after the source has been agitated in the first container 102.

As described above with respect to FIGS. 1 and 2, the second container 104 and the third container 106 can be adapted to enrich one or more microorganisms in series, or in parallel, and the second container 104 can enrich the same or a different microorganism(s) than the third container 106. For example, in some embodiments, following the method illustrated in FIG. 1, the source can be agitated in the first container 102 (e.g., by paddle blending) to form a liquid composition, at least a portion of the liquid composition can be transferred from the first container 102 to the second container 104, and the second container 104 can be adapted to enrich a microorganism of interest in the liquid composition to form a first enriched liquid composition. At least a portion of the first enriched liquid composition can be transferred from the second container 104 to the third container 106, in series, to further enrich the same or a different microorganism of interest to form a second enriched liquid composition. In some embodiments, the third container 106 is adapted to further enrich the same microorganism of interest, such that the third container 106 provides a second enrichment step to improve the efficiency and efficacy of the enrichment process. In some embodiments, the source can be agitated in the first container 102 to form a liquid composition, and at least a portion of the liquid composition can be transferred from the first container 102 into both the second container 104 and the third container 106, such that the second container 104 and the third container

106 are adapted to enrich, in parallel, one or more microorganisms of interest, where the second container 104 and the third container 106 can be adapted to enrich the same or different microorganism(s) of interest.

The second container 104 and the third container 106 can be positioned internally or externally with respect to the first container 102. In embodiments in which the second container 104 and the third container 106 are positioned internally with respect to the first container 102, contamination can be reduced by employing second and third containers

104 and 106 that are not removably coupled to the first container 102, such that the second and third containers 104 and 106 are coupled to the first container 102 from the beginning of the process (e.g., adding the source to the first container 102) and throughout enrichment, incubation, etc. Such embodiments can be useful, for example, when the enrichment steps in the second container 104 and the third container 106 are to be performed under the same enrichment conditions, such as incubation temperature, pressure, humidity, etc. In addition, in embodiments employing internal second and third containers 104, 106, the second and third containers 104 and 106 can still include the second port 124 and the third port 126, respectively, to allow material to be transferred between the first container 102, the second container 104 and the third container 106. For example, the second and third containers 104 and 106 can be coupled to an inner surface or inner wall of the first container 102 to facilitate access to the interior of the second and third containers 104 and 106.

The first container 102, the second container 104 and the third container 106 can all be formed of the same or different materials. For example, the first container 102 can be formed of a collapsible material, such that the source can be agitated in the first container 102 by means such as paddle blending. In some embodiments, the first container 102 is formed of a material that is not self-supporting. In some embodiments (including embodiments in which the first container 102 is collapsible), however, the first container 102 can be adapted to be freestanding, such as by virtue of the container design, examples of which are illustrated in FIGS. 5-10 and described below. In some embodiments, the second container 104 and/or the third container 106 can be formed of a collapsible (i.e., not self-supporting) and/or deformable material, or one or both of the second container 104 and the third container 106 can be formed of a rigid material (i.e., a material that is more rigid than the first container 102) and/or a self-supporting material. In addition, each container 102, 104, 106 does not need to be formed of the same material in its entirety. For example, the port 122, 124, 126 can be formed of a rigid and/or self- supporting material even if the majority of the respective container material is collapsible.

Two secondary containers (i.e., the second container 104 and the third container

106) are shown by way of example only, but it should be understood that the sample preparation system 100 can include fewer or more secondary containers, as necessary, which can be used in series, in parallel, or a combination thereof.

In addition, by way of example only, the first container 102 is shown in FIG. 3 as being the largest of the three containers. Such embodiments can be useful, for example, when smaller volumes can be for subsequent processing (e.g., enriching, incubating, etc.). However, it should be understood that the relative sizes and volumes are shown for the purpose of illustration and other relative sizes and volumes can be employed without departing from the spirit and scope of the present disclosure. Similarly, the shapes of the containers 102, 104 and 106 are shown by way of example only, and it should be understood that other shapes and configurations can be employed.

In some embodiments, as shown in FIG. 3, the first container 102 can include a first closed end 132 and a second open end 134. In such embodiments, the second open end 134 can be closed via one or more closures or closure means 136, which can include one or both of temporary and permanent/semi-permanent closure means. For example, in the embodiment illustrated in FIG. 3, the first container 102 includes a first temporary closure 138 and a second permanent (or semi-permanent) closure 142.

The temporary closure 138 can include a variety of temporary coupling means, including, but not limited to, a zipper (e.g., a plastic zipper, a metal zipper, a tongue-and- groove zipper, etc.); a clamp (e.g., a spring-loaded clamp, a snap-type clamp, etc.); a clip

(e.g., a spring-loaded clip, etc.); ties (e.g., wire ties); one or more magnets; tape; an adhesive; a cohesive; a hook-and-loop fastener; snap-fit engagement; press-fit engagement (also sometimes referred to as "friction-fit engagement" or "interference-fit engagement"); thermal bonding (e.g., heat and/or pressure applied to one or both of the components to be coupled); other suitable temporary coupling means; and combinations thereof. The permanent/semi-permanent closure 142 can include a variety of permanent or semipermanent coupling means, including, but not limited to, an adhesive; a cohesive; tape; stitches; staples; crimps; ties (e.g., wire ties); welding (e.g., sonic (e.g., ultrasonic) welding); thermal bonding; snap-fit engagement; press-fit engagement; heat sealing; other suitable permanent or semi-permanent coupling means; and combinations thereof. One of ordinary skill in the art will recognize that some of the permanent or semi-permanent coupling means can also be adapted to be temporary, and vice versa, and are categorized in this way by way of example only.

In embodiments employing two or more closures 136, such as the embodiment illustrated in FIG. 3, the first temporary closure 138 can be used (e.g., iteratively) in preparing the source, such as prior to agitating or further processing. For example, in some embodiments, a variety of sources (and, optionally, diluent(s)) can be combined in the first container 102. In embodiments in which the multiple sources are added (e.g., sequentially) to the first container 102, the first container 102 can be temporarily closed between successive additions of sources (and/or diluent(s)). After the sources (and diluent(s)) have been added to the first container 102, the first container 102 can be permanently or semi-permanently closed via the second permanent/semi-permanent closure 142, and the sample preparation system 100 can be used to prepare and/or process a sample. The first and second closures 138 and 142 are illustrated in FIG. 3 as being spaced a vertical distance apart, however, this relative positioning is shown for illustration purposes only. Some embodiments of the present disclosure include only one type of closure 136. For example, in some embodiments, the sample preparation system 100 includes only the permanent/semi-permanent closure 142. Still, other suitable closure configurations can be employed without departing from the spirit and scope of the present disclosure.

In the embodiment illustrated in FIG. 3, the first end 132 is closed and the second end 134 is open and adapted to be closed with the one or more closures 136. However, it should be understood that the first end 132 can also be open, and both ends 132, 134 can be closed when desired by one or more closures 136.

In use, one or more sources (and diluent(s)) can be added to the first container 102.

As mentioned above, the first temporary closure 138 can be employed as needed during this process. After the source(s) (and diluent(s)) have been added to the first container 102, the permanent/semi-permanent closure 142 can be closed to seal the contents of the first container 102 from ambience, except for any communication that may be present via the port 122 or the material makeup of the first container 102 itself. For example, in some embodiments, the first container 102 can be gas permeable and liquid impermeable. Such gas permeability can be useful, for example, if gas exchange with ambience is necessary (e.g., for the microorganism of interest).

After the closure 142 has been closed, the source can be agitated in the first container 102 to form a liquid composition. The source can be agitated within the first container 102 by any of the above-described agitation means. For example, the sample preparation system 100 can be positioned inside a paddle blender, and the collapsible nature of the first container 102 can allow the source to be agitated to form a liquid composition. If the second and third containers 104 and 106 are not already coupled to the first container 102, the second and/or third containers 104 and 106 can be coupled to the first container 102 after the source is agitated. A sample (i.e., all or a portion) of the liquid composition can be removed from the first container 102 via the port 122 and transferred to one or more of the secondary containers (i.e., the second and third containers 104 and 106). The secondary container 104, 106 can be equipped with one or more enrichment media or enrichment media can be added to the secondary container 104, 106 via the respective port 124, 126 before or after addition of the liquid composition to the secondary container 104, 106. In some embodiments, a first stage of enrichment can occur in the first container 102. Enrichment media can include any media necessary for the enrichment of one or more microorganisms of interest in the source, and can include, but are not limited to, growth media; growth inhibitor(s); any of the media listed above with respect to the diluent; other media which may provide a more favorable growth environment for the analyte(s) of interest, such as media that promotes an optimal pH, environmental gas compositions, etc.; other suitable media; and combinations thereof.

In some embodiments, enrichment media can be provided in wet or dry form. For example, in some embodiments the secondary container 104, 106 can include enrichment media in dry (e.g., powdered) form that is dispersed and/or dissolved when the liquid composition is added to the secondary container 104, 106. In some embodiments, the enrichment media is adsorbed to an inner surface of the secondary container 104, 106 and is liberated, dispersed and/or dissolved when the liquid composition is added to the secondary container 104, 106. In some embodiments, the enrichment media is provided in the form of a liquid and can be added before, during, or after addition of the liquid composition to the secondary container 104, 106. In some embodiments, the enrichment media can form a portion of a diluent that is added to the source or liquid composition in the first container 102, and the actual enrichment process (e.g., including any necessary incubation steps, etc.) can occur in one or more of the second container 104 and third container 106. In some embodiments, additional agitation can occur in one or more of the secondary containers 104 and 106, for example, to ensure adequate exposure of the source to the enrichment media.

In some embodiments, the liquid composition can be filtered at any point in the process (e.g., prior to being transferred to a secondary container 104, 106) to remove insoluble matter. For example, in some embodiments, one or more of the containers 102, 104 and 106 can be equipped with a filter. Such a filter can be positioned, for example, in fluid communication with the port 122, such that the liquid composition is filtered upon movement of the liquid composition into and out of the port 122. In some embodiments, the port 124, 126 of the secondary container 104, 106, respectively, can function as an inlet port or an outlet port, and the secondary container 104, 106 can include an additional port to function as an outlet port or an inlet port, respectively. In such embodiments, the outlet port can be equipped with a filter, such that the liquid composition (e.g., enriched liquid composition) can be filtered upon removal from the secondary container 104, 106.

In some embodiments, the secondary container 104, 106 can be adapted for incubation. For example, the secondary container 104, 106 can be formed of a material that can withstand incubation conditions for the microorganism(s) of interest, such as heat, pressure, humidity, etc. In some embodiments, one or more of the secondary container(s) 104, 106 can be removed from being coupled to the first container 102 for enrichment. In some embodiments, the first container 102 is also adapted for incubation, and the secondary container 104, 106 can remain coupled to the first container 102 during incubation and/or other processing steps. However, embodiments in which one or more of the secondary containers 104 and 106 can be removed from the first container 102 can be useful to reduce the total size and volume of material that needs to be handled and managed in subsequent processing steps.

One or more of the first container 102, the second container 104 and the third container 106 can be coupled to a detection device in order to test for the microorganism(s) of interest and/or an analyte that may be indicative of one or more of the presence, quantity, and/or viability of the microorganism(s) of interest. FIG. 4 illustrates another sample preparation system 200 according to the present disclosure, wherein like numerals represent like elements. The sample preparation system 200 shares many of the same elements and features described above with reference to the illustrated embodiment of FIG. 3. Accordingly, elements and features corresponding to elements and features in the illustrated embodiment of FIG. 3 are provided with the same reference numerals in the 200 series. Reference is made to the description above accompanying FIG. 3 for a more complete description of the features and elements (and alternatives to such features and elements) of the embodiment illustrated in FIG. 4.

The sample preparation system 200 includes a first container 202, a second container 204, and a third container 206. The first container 202 includes a first closed end 232, a second open end 234, and one or more closures 236 adapted to close the open end 234.

In the embodiment illustrated in FIG. 4, the first container 202 can be fluidly coupled to the second container 204 and the third container 206, and the second container

204 can be fluidly coupled to the third container 206. As such, a source can be transferred from the first container 202 to one or both of the second container 204 and the third container 206, from the second container 204 to the third container 206, from the third container 206 to the second container 204, or combinations thereof. Such fluid communication between the first container 202 and the second container 204 is represented schematically in FIG. 4 by a first connector 242. Similarly, fluid communication between the first container 202 and the third container 206 is represented by a second connector 244. In addition, fluid communication between the second container 204 and the third container 206 is represented by a third connector 246. Each connector 242, 244, 246 can include one or more of an aperture, a tube, and the like, and combinations thereof.

In some embodiments, the secondary container 204, 206 can be physically and fluidly coupled to the first container 202. For example, in some embodiments, the fluid communication between the first container 202 and the secondary container 204, 206 can be provided when the physical coupling is provided. That is, in some embodiments, the connector 242, 244 can be adapted to provide physical and fluid coupling (e.g., the connector 242, 244 can include a snap-fit engagement that provides physical coupling as well as fluid coupling between the respective containers). In some embodiments, the physical coupling can be separate from the fluid coupling, and the two types of coupling need not occur exactly simultaneously. For example, in some embodiments, the secondary container 204, 206 can be fluidly connected to the first container 202 via the connector 242, 244, and then, if desired, the secondary container 204, 206 can be physically coupled to the first container 202 (e.g., via a variety of coupling means, including any of the above-described coupling means, such as a hook-and-loop fastener). The fluid communication and physical coupling between the second container 204 and the third container 206 can also include any of those described above.

The second container 204 and the third container 206 can both be physically coupled to the first container 202, such that even if the second container 204 and the third container 206 are not physically coupled to each other, they can both be "anchored" with respect to the first container 202.

As described above with respect to the sample preparation system 100 illustrated in FIG. 3, the second container 204 and the third container 206 can be positioned internally or externally with respect to the first container 202, or a combination thereof (e.g., the second container 204 can be positioned internally and the third container 206 can be positioned externally, or vice versa). For example, in some embodiments, the secondary container 204, 206 can be coupled to an external portion of the first container 202, and in some embodiments, the secondary container 204 can be coupled to an internal portion (e.g., an inner surface) of the first container 202. In embodiments in which the secondary container 204, 206 is coupled to an internal portion of the first container 202, the connector 242, 244 can be positioned to provide fluid communication between the interior of the secondary container 204, 206 and the interior of the first container 202.

The first container 202 can be in fluid communication with one or more of the secondary containers 204, 206, such that a fluid path 248 is defined at least partially by the first container 202 and the respective one or more secondary containers 204, 206. The fluid path 248 can allow at least a portion of a source, a liquid composition, an enriched liquid composition, and/or a filtrate of any of the above, to be moved between the first container 202 and one or more of the secondary containers 204, 206 by moving in the fluid path 248 and not being exposed to ambience during the transfer. Similarly, in some embodiments, such as the embodiment illustrated in FIG. 4, two or more of the secondary containers 204, 206 can be in fluid communication, such that the fluid path 248 is further defined at least partially by the additional secondary container(s) 204, 206. In such embodiments, the fluid path 248 can further allow at least a portion of a source, a liquid composition, an enriched liquid composition, and/or a filtrate of any of the above, to be moved between the one or more secondary containers 204, 206 by moving in the fluid path 248 and not being exposed to ambience during the transfer.

In some embodiments, the phrase "without exposing to ambience" and derivations thereof refers to not removing a sample (i.e., at least a portion of a source, a liquid composition, an enriched liquid composition, and/or a filtrate of any of the above, etc.) from the sample preparation system 200 during the transfer between the first container 202 and one or more of the secondary containers 204, 206 (e.g., to prevent spills or contamination), such that the sample remains in the fluid path 248 of the sample preparation system 200 from agitation to enrichment, or even to another downstream step, but does not necessarily mean that the sample preparation system 200 is closed to gas- exchange or that other liquids cannot be introduced into the sample preparation system 200. For example, in some embodiments, a container 202, 204, 206, or a portion thereof, is gas-permeable, or includes a gas-permeable film or membrane (e.g., for aerobic bacteria to continue to have access to oxygen).

As with the sample preparation system 100 illustrated in FIG. 3 and described above, the sample preparation system 200 can allow for serial and/or parallel enrichment. For example, the material transfer between the containers 202, 204 and 206 can be controlled, such that a liquid composition can be moved only from the first container 202 to one of the secondary containers 204 and 206 for a first enrichment stage, and then moved from the first secondary container 204/206 to a second secondary container 204/206 for a second enrichment stage (i.e., serial enrichment). In some embodiments, the material transfer between the containers 202, 204 can be controlled, such that a liquid composition can be moved from the first container 202 to each of the secondary containers 204 and 206 for enrichment (i.e., parallel enrichment). As described above, a combination of serial and parallel enrichment can be employed, and additional secondary containers can be used for additional enrichment and/or downstream processing steps. Two secondary containers 204, 206 are shown by way of example only.

Such control of material transfer between containers 202, 204 and 206 can be performed in a variety of ways, including positioning one or more valves in the fluid path 248. For example, a first valve 252 is illustrated schematically in FIG. 4 as being positioned in the first connector 242 to control movement of a liquid composition between the first container 202 and the second container 204. Similarly, a second valve 254 is illustrated in FIG. 4 as being positioned in the second connector 244, and a third valve 256 is illustrated in FIG. 4 as being positioned in the third connector 246. Each valve 252, 254, 256 can be adapted to change between a first (closed) state in which the upstream container (e.g., the first container 202) and the downstream container (e.g., the second container 204) are not in fluid communication and a second (open) state in which the upstream container and the downstream container are in fluid communication. Each valve 252, 254, 256 can be actuated to change between the first state and the second state, depending on what type of valve is employed.

Suitable valves 252, 254 and 256 can include, but are not limited to, a variety of manual or automatic valves, an electronic pressure transducer, check valves (e.g., duckbill valves, ball check valves, diaphragm check valves, swing check valves, stop check valves, lift check valves, etc.) or other types of valves, such as stopcock valves, butterfly valves, metering valves, constant volume metering valves, timer valves, other one-way valves, other suitable valves, and combinations thereof.

For example, in some embodiments, one or more of the valves 252, 254 and 256 can include a one-way pressure-activated or pressure-controlled valve. The first valve 252 will now be described by way of example only, for simplicity, but it should be understood that the description can be applied to the other valves 254 and 256 as well.

The valve 252 can be positioned in fluid communication between the first container 202 and the second container 204. In some embodiments, the valve 252 can be adapted to allow a liquid composition to move from the first container 202 to the second container 204 when a sufficient pressure differential is established between the first container 202 and the second container 204. For example, the valve 252 can be actuated to open by applying positive pressure to the first container 202 and/or by applying negative pressure to the second container 204 (i.e., to the downstream side of the valve 252 when a one-way valve is employed). Negative pressure can be applied to the second container 204 in a variety of ways, including, but not limited to, by compressing (e.g., manually or automatically) the second container 204 and releasing it and/or by coupling a vacuum source to the valve 252, for example, by coupling a vacuum source to the second container 204. A vacuum source can include, but is not limited to, a mechanical pump that creates a reduced pressure, or a manual pump (e.g., a syringe-plunger combination), and combinations thereof.

In some embodiments, the pressure differential for opening the valve 252 can be established by the mass of liquid on the upstream side of the valve 252. For example, a liquid composition can cause sufficient pressure (e.g., head pressure when the sample preparation system 200 is tipped or inverted) to open the valve 252.

When a sufficient pressure differential is established, the valve 252 can control how a liquid composition is dispensed into the second container 204. For example, depending on the type of valve 252 used, the liquid composition can be dispensed into the second container 204 in a continuous stream, in a drop-wise fashion, or in another suitable flow configuration. Such control can allow the valve to dispense at a desired volumetric flow rate or volumetric metering scheme to achieve a desired volume of the liquid composition in the second container 204.

In embodiments employing such a pressure-activated valve, the first container 202, the second container 204 and the valve 252 can be adapted such that the valve 252 will not be actuated to open prematurely. For example, the first container 202, the second container 204 and/or the first valve 252 can be adapted such that compression that may occur during agitation of the source will not be sufficient to actuate the valve 252, such that the liquid composition cannot be transferred to the second container 204 prematurely. In some embodiments, the sample preparation system 200 can be adapted such that compression-based agitation can cause some of the liquid composition to be transferred to the second container 204 at a desired point during agitation. A variety of options are available, depending on the amount of initial agitation desired, the microorganism of interest, etc.

In some embodiments, the valve 252 can be formed so as not include any movable parts but rather includes a restricted opening, such as a tip that has a gradually decreasing cross-sectional area. In such embodiments, a liquid composition will not be able to pass through the restricted opening until sufficient pressure is established to force the liquid composition out of the restricted opening.

In some embodiments, the sample preparation system 200 can include one or more outgassing valves positioned anywhere in the fluid path 248 to provide fluid communication between the fluid path 248 and ambience. For example, in some embodiments, the components of the sample preparation system 200 are not gas- permeable, but if gases are developing in the sample preparation system 200 (e.g., if bacteria in the sample preparation system 200 are producing gas, or if a reaction is taking place between the source and another mixture component (e.g., enrichment media) that produces gas, or if the agitation process produces a build-up of gas) and enough pressure develops within the fluid path 248, gas can be released via an outgassing valve. Such outgassing valves can include any of the above-described valves to allow for outgassing of the sample preparation system 200.

Furthermore, in embodiments in which the components of the sample preparation system 200 are not gas-permeable, anaerobic bacteria may be positioned (and cultured) in the sample preparation system 200 by replacing the air in the sample preparation system 200 with an oxygen-free environment such as carbon dioxide. In such embodiments, the replacement gas can be introduced into the sample preparation system 200 via a valve or an inlet tube, and air can be removed from the sample preparation system 200 via an outgassing valve, and/or via a port (e.g., one of the ports 122, 124, 126 illustrated in FIG. 3). As a result, an outgassing valve can further allow for outgassing when the atmosphere in the sample preparation system 200 is replaced. In use, one or more sources (and diluent(s)) can be added to the first container 202, and the one or more closures 236 can be employed as needed, as described above with respect to the sample preparation system 100. In the embodiment illustrated in FIG. 4, the source and liquid composition need not be exposed to ambience again, due to the internal fluid coupling between containers 202, 204 and 206. After the first container 202 has been closed, the source can be agitated in the first container 202. The source can be agitated within the first container 202 by any of the above-described agitation means. If the second and third containers 204 and 206 are not already coupled to the first container 202, the second and/or third containers 204 and 206 can be coupled to the first container 202 after the source is agitated. A sample (i.e., all or a portion) of the liquid composition can be moved from the first container 202 via one or more of the first and second connectors 242 and 244 (e.g., by actuating one or both of the valves 252 and 254, respectively, to open) to one or more of the secondary containers (i.e., one or more of the second and third containers 204 and 206). The secondary container 204, 206 can be equipped with one or more enrichment media before or after addition of the liquid composition to the respective secondary container 204, 206. If serial enrichment is necessary, a first enriched liquid composition can be transferred from one secondary container 204, 206 to another secondary container 206, 204 (e.g., via the third connector 256) and so on.

The sample preparation system 200 can be provided with all of the above- described connectors 242, 244, 246 and valves 252, 254, 256, and the sample preparation system 200 can then be configured to meet a user's needs, or the various connectors 242, 244, 246 and/or valves 252, 254, 256 can be selectively employed. For example, if only a two-stage serial enrichment is desired, the second connector 244 and/or the second valve 254 can be closed or blocked (or, alternatively, only the first and third connectors 242, 246 and/or the first and third valves 252, 256 can be activated to function), such that fluid communication can only occur from the first container 202 to the second container 204, and then to the third container 206. Such control or selective activation of the connectors 242, 244, 246 and/or valves 252, 254, 256 can occur prior to using the sample preparation system 200 or during use of the sample preparation system 200. FIG. 3 illustrates a sample preparation system 100 that includes ports 122, 124, 126 that allow for physical transfer of a source between the containers 102, 104 and 106. FIG. 4 illustrates a sample preparation system 200 that includes connectors 242, 244, 246 to allow the source to be transferred between the containers 202, 204 and 206 without exposure to ambience. It should be understood that the sample preparation systems of the present disclosure can include combinations of the sample preparation systems 100 and 200, such that a combination of physical transfer and fluid communication between containers can be employed.

FIGS. 5 and 6 illustrate another sample preparation system 300 according to the present disclosure, wherein like numerals represent like elements. The sample preparation system 300 shares many of the same elements and features described above with reference to the illustrated embodiments of FIGS. 3 and 4. Accordingly, elements and features corresponding to elements and features in the illustrated embodiments of FIGS. 3 and 4 are provided with the same reference numerals in the 300 series. Reference is made to the description above accompanying FIGS. 3 and 4 for a more complete description of the features and elements (and alternatives to such features and elements) of the embodiment illustrated in FIGS. 5 and 6.

The sample preparation system 300 includes a first container 302, a second container 304, and a third container 306. The first container 302 includes a first closed end 332, a second open end 334, and one or more closures 336 adapted to close the open end 334. In some embodiments, as shown in FIGS. 5 and 6, the first container 302 can be formed in a first portion 303 of the sample preparation system 300, and the second and third containers 304 and 306 can be formed in a second portion 305 of the sample preparation system 300.

In some embodiments, the sample preparation system 300, and particularly, the first container 302, can be adapted to be freestanding. In some embodiments, one of the ends 332, 334 of the first container 302 can be adapted to be freestanding. For example, as shown in FIGS. 5 and 6, the first container 302 can include one or more walls 331 and a base 333. One or more of the walls 331 can be equipped with fold lines or the like to facilitate collapsing the first container 302 into a flat configuration for storage and/or transportation.

The base 333 can be configured (or one or more of the walls 331 and/or the base 333 can be gusseted), such that the first container 302 can stand on its own (e.g., on the first end 332), particularly when a source or liquid composition is positioned within the first container 302, even when the first container 302 is not formed of a self-supporting material (e.g., when the first container 302 is formed of a collapsible or deformable material). The base 333 of the embodiment illustrated in FIGS. 5 and 6 is structured similar to that of a shopping bag to provide the freestanding feature, such that the first container 302 can be flattened for storage and/or transportation and expanded and stood on end as desired. However, it should be understood that the first container 302, or any other component of the sample preparation system 300, can be configured to be freestanding in a variety of other ways. For example, in some embodiments, the first container 302 can include a flange, a gusset, or an extension of material around the first end 332. Such a flange can be formed of reinforced material, such that the first container 302 can stand up on such a flange, particularly when a source or liquid composition is positioned within the first container 302.

Similar to the sample preparation system 200 illustrated in FIG. 4 and described above, the sample preparation system 300 can include a fluid path 348 (see FIG. 5) that is defined at least partially by the containers 302, 304 and 306. That is, the first container 302 can be fluidly coupled to the second container 304 and the third container 306, and the second container 304 can be fluidly coupled to the third container 306. As such, a liquid composition can be transferred from the first container 302 to one or both of the second container 304 and the third container 306, from the second container 304 to the third container 306, from the third container 306 to the second container 304, or combinations thereof, as selected by a user. Such fluid communication between the first container 302 and the second container 304 is represented schematically in FIGS. 5 and 6 by a first connector 342. Similarly, fluid communication between the first container 302 and the third container 306 is represented by a second connector 344. In addition, fluid communication between the second container 304 and the third container 306 is represented by a third connector 346. In addition, a first valve 352 can be positioned in or adjacent the first connector 342, a second valve 354 can be positioned in or adjacent the second connector 344, and a third valve 356 can be positioned in or adjacent the third connector 346. A user can control and/or selectively use and/or actuate the connectors 342, 344 and 346 and/or the valves 352, 354 and 356 to achieve the desired number and arrangement of enrichment steps (e.g., serial, parallel, or a combination thereof).

By way of example only, the second portion 305 of the sample preparation system 300 is positioned internally with respect to the first container 302, and the interior of second portion 305 is separated from the interior of the first container 302 by one or more inner walls 360, which can allow for facile manufacturing. As a result, the second portion 305 can be defined by one or more walls 331 of the first container 302 (e.g., in the embodiment illustrated in FIGS. 5 and 6, the second portion 305 is defined by a first wall 331a, a second wall 331b and a third wall 331c) and an inner wall 360 positioned to separate the second portion 305 and the first portion 303. In addition, the second portion 305 can be defined by one or more seams and/or welds. For example, the second portion 305 can be defined by one or more side seams and/or welds, instead of second and third walls 331b and 331c of the first container 302. By way of example only, as shown in FIGS. 5 and 6, the second portion 305 is further defined by a first (lower) seam or weld 362 positioned toward the first end 332 of the first container 302 and a second (upper) seam or weld 364 positioned toward the second end 334 of the first container 302.

The second portion 305 of the sample preparation system 300 can include as few or as many containers within it as desired (e.g., given the desired sample preparation and processing steps) and as structurally possible. For example, as shown in FIG. 5, the second portion 305 includes the second container 304 and the third container 306, separated from each other by a third seam or weld (or wall) 366, the third connector 346 providing fluid communication across the third seam 366. Any of the seams and/or welds 362, 364, 366 can be formed by any of the coupling means described above with respect to the closures, such as thermal bonding, adhesives, and the like, and combinations thereof.

By way of example only, all of the containers 302, 304 and 306 are adapted to be fluidly coupled to one another, such that a liquid composition can be moved and enriched, in series and/or in parallel, without exposing the liquid composition to ambience after the source has been added to the first container 302. As a result, the sample preparation system 300 would function in use similarly to the sample preparation system 200 illustrated in FIG. 4 and described above. However, it should be understood that a combination of the fluid communication structures illustrated in FIGS. 5 and 6 and the ports illustrated in FIG. 3 can be employed to achieve the desired sample preparation and processing steps.

In some embodiments, the second portion 305 (or a portion thereof) can be removably coupled (e.g., via any of the above-described temporary coupling means) to the first portion 303. For example, in some embodiments, the second portion 305 can be defined by an inner wall positioned adjacent a wall 331 of the first container 302, rather than being defined directly by a wall 331 itself. In such embodiments, the first container 302 can be opened after the liquid composition has been transferred to the second portion 305, and the second portion 305 (or a portion thereof) can be removed from the first container 302 for subsequent processing (e.g., incubation). In such embodiments, contamination and sample loss can be minimized by transferring substantially all of the liquid composition into the second portion 305 prior to removing the second portion 305. While such embodiments are possible, advantageous results can be achieved by not removing the second portion 305 from the first portion 303, such as reduced risk of contamination, sample loss, and the like.

FIGS. 7 and 8 illustrate another sample preparation system 400 according to the present disclosure, wherein like numerals represent like elements. The sample preparation system 400 shares many of the same elements and features described above with reference to the illustrated embodiments of FIGS. 4-6. Accordingly, elements and features corresponding to elements and features in the illustrated embodiments of FIGS. 4-6 are provided with the same reference numerals in the 400 series. Reference is made to the description above accompanying FIGS. 4-6 for a more complete description of the features and elements (and alternatives to such features and elements) of the embodiment illustrated in FIGS. 7 and 8.

The sample preparation system 400 includes a first container 402, a second container 404, and a third container 406. The first container 402 includes a first closed end 432, a second open end 434, and one or more closures 436 adapted to close the open end 434. In some embodiments, as shown in FIGS. 7 and 8, the first container 402 can be formed in a first portion 403 of the sample preparation system 400, and the second and third containers 404 and 406 can be formed in a second portion 405 of the sample preparation system 400.

As shown in FIGS. 7 and 8, the sample preparation system 400, and particularly, the first container 402 can be adapted to be freestanding. For example, as shown in FIGS.

7 and 8, the first container 402 can include one or more walls 431 and a base 433. One or more of the walls 431 can be equipped with fold lines or the like to facilitate collapsing the first container 402 into a flat configuration for storage and/or transportation.

Similar to the sample preparation system 300 illustrated in FIGS. 5 and 6 and described above, the base 433 can be configured (or one or more of the walls 431 and/or the base 433 can be gusseted), such that the first container 402 can stand on its own (e.g., on the first end 432), particularly when a source or liquid composition is positioned within the first container 402, even when the first container 402 is not formed of a self-supporting material (e.g., when the first container 402 is formed of a collapsible or deformable material).

Similar to the sample preparation system 200 illustrated in FIG. 4 and the sample preparation system 300 illustrated in FIGS. 5 and 6, the sample preparation system 400 can include a fluid path 448 (see FIG. 7) that is defined at least partially by the containers 402, 404 and 406. That is, the first container 402 can be fluidly coupled to the second container 404 and the third container 406, and the second container 404 can be fluidly coupled to the third container 406. As such, a liquid composition can be transferred from the first container 402 to one or both of the second container 404 and the third container 406, from the second container 404 to the third container 406, from the third container 406 to the second container 404, or combinations thereof, as selected by a user. Such fluid communication between the first container 402 and the second container 404 is represented schematically in FIGS. 7 and 8 by a first connector 442. Similarly, fluid communication between the first container 402 and the third container 406 is represented by a second connector 444. In addition, fluid communication between the second container 404 and the third container 406 is represented by a third connector 446. In addition, a first valve 452 can be positioned in or adjacent the first connector 442, a second valve 454 can be positioned in or adjacent the second connector 444, and a third valve 456 can be positioned in or adjacent the third connector 446. A user can control and/or selectively use and/or actuate the connectors 442, 444 and 446 and/or the valves

452, 454 and 456 to achieve the desired number and arrangement of enrichment steps (e.g., serial, parallel, or a combination thereof).

By way of example only, the second portion 405 of the sample preparation system 400 is positioned externally with respect to the first container 402. While a variety of configurations of the second portion 405 are possible, the second portion 405 is illustrated in FIGS. 7 and 8 as being defined by a front wall 470, a rear wall 472 positioned adjacent a wall 431 of the first container 402, a left side wall 474, and a right side wall 476. The second portion 405 can be further defined by an upper wall, seam or weld, and a lower wall, seam, or weld. In the embodiment illustrated in FIGS. 7 and 8, the second portion 405 is further defined by a first (lower) seam or weld 462 positioned toward the first end

432 of the first container 402 and a second (upper) seam or weld 464 positioned toward the second end 434 of the first container 402.

The second portion 405 can include as few or as many containers within it as desired (e.g., given the desired sample preparation and processing steps) and as structurally possible. For example, as shown in FIG. 7, the second portion 405 includes the second container 404 and the third container 406, separated from each other by a third seam or weld (or wall) 466, the third connector 446 providing fluid communication across the third seam 466. Any of the seams and/or welds 462, 464, 466 can be formed by any of the coupling means described above with respect to the closures, such as thermal bonding, adhesives, and the like, and combinations thereof.

By way of example only, all of the containers 402, 404 and 406 are adapted to be fluidly coupled to one another, such that a liquid composition can be moved and enriched, in series and/or in parallel, without exposing the liquid composition to ambience after the source has been added to the first container 402. As a result, the sample preparation system 400 would function in use similarly to the sample preparation system 200 illustrated in FIG. 4 and described above. However, it should be understood that a combination of the fluid communication structures illustrated in FIGS. 7 and 8 and the ports illustrated in FIG. 3 can be employed to achieve the desired sample preparation and processing steps.

In the embodiment illustrated in FIGS. 7 and 8, the second portion 405 is adapted to be removable from the first portion 403 (e.g., for further processing, such as incubation, etc.). As a result, any portion of the rear wall 472 can be coupled to the first portion 403 (e.g., the end of the second portion 405 adjacent the second seam 464 can be coupled to a wall 431 of the first container 402) via any of the above-described temporary coupling means, such as a hook-and-loop fastener, a zipper, and the like, and combinations thereof. Alternatively, or in addition, one or more of the first and second connectors 442 and 444 can be adapted to provide physical coupling in addition to fluid coupling, as described above, such as a snap-fit engagement, or the like. In such removable embodiments, the second portion 405 can be removed from the first portion 403 after the liquid composition has been transferred to one or more of the second container 404 and the third container

406, and the first portion 403 can be discarded.

In addition, in such embodiments, the second portion 405 can include sealing means to prevent leakage from the second and third containers 404 and 406 when the second portion 405 is removed from the first portion 403. Alternatively, or in addition, the first and second valves 452 and 454 can be adapted to remain closed after the second portion 405 is removed from the first portion 403, until desired. Alternatively, or in addition, the first and second connectors 442 and 444 can each be equipped with two or more valves 452 and 454, such that the first container 402, the second container 404 and the third container 406 will remain closed and sealed, until desired, when the second portion 405 is removed from the first portion 403. A variety of other sealing mechanisms, valves and/or closures can be employed to selectively close and provide suitable sealing of the containers 402, 404 and 406 when the second portion 405 is removed from the first portion 403.

In some embodiments, the second portion 405 can be defined at least partially by the first portion 403 (e.g., a wall 431 of the first container 402), such that the second portion 405 is not removably coupled to the first portion 403, but rather the entire sample preparation system 400 can be processed (e.g., incubated) at a time. For example, in some embodiments, the second portion 405 can be defined by a wall 431 of the container 402, the front wall 470, and upper, lower, left and right walls, seams and/or welds. Such embodiments can allow for facile manufacturing processes.

FIGS. 9 and 10 illustrate another sample preparation system 500 according to the present disclosure, wherein like numerals represent like elements. The sample preparation system 500 shares many of the same elements and features described above with reference to the illustrated embodiments of FIGS. 4-8. Accordingly, elements and features corresponding to elements and features in the illustrated embodiments of FIGS. 4-8 are provided with the same reference numerals in the 500 series. Reference is made to the description above accompanying FIGS. 4-8 for a more complete description of the features and elements (and alternatives to such features and elements) of the embodiment illustrated in FIGS. 9 and 10.

The sample preparation system 500 includes a first container 502, a second container 504, and a third container 506. The sample preparation system 500 includes a first closed end 532, a second closed end 537, an opening 534 (see FIG. 9), and one or more closures 536. In some embodiments, as shown in FIGS. 9 and 10, the first container 502 can be formed in a first portion 503 of the sample preparation system 500 (e.g., adjacent the first end 532), and the second and third containers 504 and 506 can be formed in a second portion 505 of the sample preparation system 500 (e.g., adjacent the second end 537). FIG. 9 illustrates the sample preparation system 500 in a first extended configuration, and FIG. 10 illustrates the sample preparation system 500 in a second folded configuration.

As shown in FIGS. 9 and 10, the sample preparation system 500, and particularly, the first container 502, can be adapted to be freestanding. For example, as shown in FIGS. 9 and 10, the first container 502 can include one or more walls 531 and a base 533. One or more of the walls 531 can be equipped with fold lines or the like to facilitate collapsing the first container 402 into a flat configuration for storage and/or transportation. Similar to the sample preparation system 300 illustrated in FIGS. 5 and 6 and described above, the base 533 can be configured (or one or more of the walls 531 and/or the base 533 can be gusseted), such that the first container 502 can stand on its own (e.g., on the first end 532), particularly when a source or liquid composition is positioned within the first container 502, even when the first container 502 is not formed of a self-supporting material (e.g., when the first container 502 is formed of a collapsible or deformable material).

Similar to the sample preparation systems 200, 300 and 400, the sample preparation system 500 can include a fluid path 548 (see FIG. 9) that is defined at least partially by the containers 502, 504 and 506. That is, the first container 502 can be fluidly coupled to the second container 504 and the third container 506, and the second container 504 can be fluidly coupled to the third container 506. As such, a liquid composition can be transferred from the first container 502 to one or both of the second container 504 and the third container 506, from the second container 504 to the third container 506, from the third container 506 to the second container 504, or combinations thereof, as selected by a user. Such fluid communication between the first container 502 and the second container 504 is represented schematically in FIGS. 9 and 10 by a first connector 542. Similarly, fluid communication between the first container 502 and the third container 506 is represented by a second connector 544. In addition, fluid communication between the second container 504 and the third container 506 is represented by a third connector 546. In addition, a first valve 552 can be positioned in or adjacent the first connector 542, a second valve 554 can be positioned in or adjacent the second connector 544, and a third valve 556 can be positioned in or adjacent the third connector 546. A user can control and/or selectively use and/or actuate the connectors 542, 544 and 546 and/or the valves 552, 554 and 556 to achieve the desired number and arrangement of enrichment steps

(e.g., serial, parallel, or a combination thereof).

In some embodiments, as shown in FIGS. 9 and 10, the first portion 503 and the second portion 505 can be formed at opposite ends of the sample preparation system 500.

In such embodiments, the second portion 505 (and the second container 504 and the third container 506) can be at least partially defined by the same walls 531 that define the first portion 503 (and the first container 502). As shown in FIGS. 9 and 10, the sample preparation system 500 includes a front wall 531a, a rear wall 531b, a left side wall 531c, and a right side wall 53 Id. Each of the walls 531a, 531b, 531c and 53 Id define the first container 502 in the first portion 503 at one end of the sample preparation system 500 and the second and third containers 504 and 506 in the second portion 505 at an opposite end of the sample preparation system 500. The first portion 503 is further defined by the base

533, and the second portion 505 is further defined by the second closed end 537.

The second portion 505 of the sample preparation system 500 can include as few or as many containers within it as desired (e.g., given the desired sample preparation and processing steps) and as structurally possible. For example, as shown in FIG. 9, the second portion 505 includes the second container 504 and the third container 506, separated from each other by a seam or weld (or wall) 566, the third connector 546 providing fluid communication across the seam 566. The seam 566 can be formed by any of the coupling means described above with respect to the closures, such as thermal bonding, adhesives, and the like, and combinations thereof.

By way of example only, all of the containers 502, 504 and 506 are adapted to be fluidly coupled to one another, such that a liquid composition can be moved and enriched, in series and/or in parallel, without exposing the liquid composition to ambience after the source has been added to the first container 502. As a result, the sample preparation system 500 would function in use similarly to the sample preparation system 200 illustrated in FIG. 4 and described above. However, it should be understood that a combination of the fluid communication structures illustrated in FIGS. 9 and 10 and the ports illustrated in FIG. 3 can be employed to achieve the desired sample preparation and processing steps.

In use, a source can be added to the first container 502 via the opening 534. For example, as shown in FIG. 10, the second portion 505 can be diverted to the side (e.g., allowed to hang adjacent the first portion 503) to facilitate accessing the opening 534. With reference to FIG. 9, after the source is added to the first container 502, a closure 538 (see FIG. 9) can be used to close the opening 534. For example, in some embodiments, the opening 534 is in one wall 531, such as the front wall 531a. By way of example only, a temporary closure 538 can be used to secure the front wall 531a to the rear wall 531b after the source has been added to the first container 502. Such a temporary closure 538 can include any of the above-described temporary coupling means. Such a temporary closure 538 can be configured to withstand the pressures generated in the first container 502 as a result of agitating the source in the first container

502. Alternatively, or in addition, the second portion 505 can be at least partially defined by one or more seams and/or welds 562 adapted to withstand the agitation process. In such embodiments, the opening 534 can be permanently or semi-permanently closed (e.g., by heat sealing) after the source has been added. Alternatively, or in addition, the sample preparation system 500 can be positioned in an agitation device, such as a paddle blender, in a way that the second portion 505 is allowed to hang outside of the paddle blender, the paddle blender providing the temporary closure 538, if necessary (e.g., by clamping the sample preparation system 500 in the paddle blender at the closure line 538).

After the source has been agitated, the temporary closure 538 can be removed or opened, and, if necessary, the opening 534 can be permanently or semi-permanently closed (e.g., by heat sealing). The liquid composition can be transferred to the second portion 505, and particularly, can be selectively transferred to one or more of the second container 504 and the third container 506 (e.g., via one or more of the connectors 542,

544, 546 and/or one or more of the valves 552, 554, 556). After the desired amount of the liquid composition has been transferred to one or more of the second container 504 and the third container 506, the second portion 505 can be closed or fluidly decoupled from the first portion 503 via a permanent or semi-permanent closure 542, for example, using any of the above-described permanent or semi-permanent coupling means. That is, the front wall 531a can be secured to the rear wall 53 Ib at the closure line 542 to permanently or semi-permanently seal the second portion 505 (and the secondary containers 504 and/or

506) from the first portion 503 (and the first container 502).

Then, the entire sample preparation system 500 can be used in subsequent processing steps, or one or more of the secondary containers 504 and 506 can be removed from the first container 502 for subsequent processing steps, for example, by removing the second portion 505 from the first portion 503. For example, in some embodiments, the second portion 505 can be separated from the first portion 503 along a perforation line 580, and the second portion 505 (or a portion thereof) can be used in subsequent processing steps.

The opening 534, the temporary closure 538, the permanent/semi-permanent closure 542, and the perforation line 580 are shown in their relative positions in FIGS. 9 and 10 solely for illustration purposes. However, it should be understood that other positions can be employed, including overlapping positions. For example, the opening 534 can instead be positioned below the temporary closure 538, the temporary closure 538 can be positioned above the perforation line 580, the temporary closure 538 and the permanent/semi-permanent closure 542 can be located at the same vertical position, etc.

FIG. 11 illustrates another sample preparation system 600 according to the present disclosure, wherein like numerals represent like elements. The sample preparation system 600 shares many of the same elements and features described above with reference to the illustrated embodiment of FIGS. 3-10. Accordingly, elements and features corresponding to elements and features in the illustrated embodiments of FIGS. 3-10 are provided with the same reference numerals in the 600 series. Reference is made to the description above accompanying FIGS. 3-10 for a more complete description of the features and elements (and alternatives to such features and elements) of the embodiment illustrated in FIG. 11.

The sample preparation system 600 includes a first container 602 and a second container 604. The sample preparation system 600 includes one "secondary" container

(i.e., the second container 604) by way of example only, but it should be understood that additional secondary containers (i.e., in a serial and/or parallel arrangement) can be similarly employed without departing from the spirit and scope of the present disclosure.

The sample preparation system 600 includes a first closed end 632, a second open end 634, and one or more closures 636.

In the embodiment illustrated in FIG. 11, the first container 602 includes a port 622, and a barrier 623 is positioned in or over the port 622. While the first container 602 is collapsible and deformable to allow for sufficient agitation of the source in the first container 602, the second container 604 is rigid (i.e., more rigid than the first container 602) and self-supporting. In some embodiments, the second container 604 can also be deformable, but the second container 604 is not collapsible. In some embodiments, the second container 604 can be evacuated to facilitate movement of a liquid composition from the first container 602 to the second container 604.

The sample preparation system 600 can further include a connector 642 dimensioned to be received in the port 622 to provide fluid communication between the first container 602 and the second container 604 when necessary. As such, the connector 642 can provide physical coupling as well as fluid coupling between the first and second containers 602 and 604. By way of example only, the connector 642 is shown as forming a portion of the second container 604. However, it should be understood that other types of connectors, physical coupling means and fluid coupling means can be employed without departing from the spirit and scope of the present disclosure. As with the embodiments described above, the second container 604 can be coupled to the first container 602 before or after the source is agitated. The port 622 and/or the barrier 623 can be adapted (e.g., equipped with one or more valves) to inhibit leakage from the first container 602 or premature transfer of the liquid composition.

The sample preparation system 600 can include a fluid path 648 defined at least partially by the first container 602, and additionally defined by the second container 604 (and the connector 642, when employed) when the second container is coupled to the first container 602. In some embodiments, a valve 652 can be positioned in the fluid path 648 to control the movement of a liquid composition from the first container 602 to the second container 604. For example, as shown in FIG. 11, the valve 652 can be positioned in or adjacent the connector 642 to control and/or meter a liquid composition into the second container 604.

After the liquid composition has been transferred to the second container 604, the second container 604 can be decoupled from the first container 602, and the second container 604 can be used in subsequent processing steps (e.g., incubation). One advantage of the rigid, self-supporting second container 604 is that the second container 604 can be easily transported, handled and/or stored (e.g., stacked) after being removed from the first container 602. In addition, in some embodiments, as shown in FIG. 11, the second container 604 can include one or more indicia 682 to facilitate transferring a desired volume of the source from the first container 602 to the second container 604.

In some embodiments, the sample preparation system 600 can include additional secondary containers (not shown) for subsequent processing steps (e.g., additional enrichment steps) that can be coupled to the second container 604 (e.g., via the connector 642) for additional serial and/or parallel enrichment.

In some embodiments, all or portion of any of the above-described sample preparation systems 100, 200, 300, 400, 500 and 600 can be disposable.

The embodiments described above and illustrated in the figures are presented by way of example only and are not intended as a limitation upon the concepts and principles of the present disclosure. As such, it will be appreciated by one having ordinary skill in the art that various changes in the elements and their configuration and arrangement (particularly relevant to mechanical cases) are possible without departing from the spirit and scope of the present disclosure. In addition, any combination of the above-described sample preparation systems 100, 200, 300, 400, 500 and 600 are possible and within the spirit and scope of the present disclosure. Various features and aspects of the present disclosure are set forth in the following claims.

Claims

WHAT IS CLAIMED IS:

1. A sample preparation system comprising: a first container adapted to contain a liquid composition comprising a source, the first container being collapsible and adapted to allow the liquid composition to be agitated; and a second container adapted to be coupled to the first container, the second container adapted to receive at least a portion of the liquid composition from the first container, and further adapted to enrich a microorganism of interest in the liquid composition to form an enriched liquid composition.

2. A sample preparation system comprising: a first container containing a liquid composition comprising a source, the first container being collapsible; and a second container coupled to the first container, the second container containing at least a portion of the liquid composition, the second container adapted to enrich a microorganism of interest in the liquid composition to form an enriched liquid composition.

3. The sample preparation system of claim 1, wherein the second container is adapted to be positioned internally with respect to the first container.

4. The sample preparation system of claim 1 or claim 3, wherein the second container is adapted to be positioned externally with respect to the first container.

5. The sample preparation system of any one of claims 1 and 3-4, wherein the second container is adapted to be in fluid communication with the first container.

6. The sample preparation system of claim 2, wherein the first container is in fluid communication with the second container.

7. The sample preparation system of any one of claims 1 and 3-5, further comprising a valve positioned between the first container and the second container when the first container is coupled to the second container, the valve configured to change between a first state in which the first container and the second container are in fluid communication and a second state in which the first container and the second container are not in fluid communication.

8. The sample preparation system of claim 2 or claim 6, wherein the first container and the second container each define at least a portion of a fluid path, and wherein a valve is positioned in the fluid path between the first container and the second container.

9. The sample preparation system of claim 8, wherein the valve is configured to change between a first state in which the first container and the second container are in fluid communication and a second state in which the first container and the second container are not in fluid communication.

10. The sample preparation system of any one of claims 7-9, wherein the valve includes a one-way pressure-activated valve.

11. The sample preparation system of any one of claims 2 and 8-10, wherein the first container and the second container are integrally formed.

12. The sample preparation system of any one of claims 2 and 8-11, wherein the second container is coupled to a detection system.

13. The sample preparation system of any one of claims 1, 3-5, 7 and 10, further comprising a third container adapted to be coupled to at least one of the first container and the second container, the third container being adapted to receive at least a portion of at least one of the liquid composition and the enriched liquid composition from at least one of the first container and the second container.

14. The sample preparation system of claim 13, wherein the third container is adapted to be in fluid communication with at least one of the first container and the second container.

15. The sample preparation system of any one of claims 2 and 8-12, further comprising a third container coupled to at least one of the first container and the second container, the third container containing at least a portion of at least one of the liquid composition and the enriched liquid composition from at least one of the first container and the second container.

16. The sample preparation system of claim 15, wherein the third container is in fluid communication with at least one of the first container and the second container.

17. The sample preparation system of any one of claims 13-16, wherein the second container and the third container are arranged in series.

18. The sample preparation system of any one of claims 13-16, wherein the second container and third container are arranged in parallel.

19. The sample preparation system of any one of claims 13-18, wherein the third container is adapted to enrich the microorganism of interest in at least one of the liquid composition and the enriched liquid composition.

20. The sample preparation system of any one of claims 13-19, wherein the microorganism of interest is a first microorganism of interest, wherein the third container is adapted to enrich a second microorganism of interest in at least one of the liquid composition and the enriched liquid composition, and wherein the second microorganism of interest is different from the first microorganism of interest.

21. The sample preparation system of any one of claims 13-20, wherein the third container is adapted for incubation.

22. The sample preparation system of any one of claims 13-21, wherein the third container is collapsible.

23. The sample preparation system of any one of claims 13-22, wherein the first container is positioned in a first portion of the sample preparation system, and wherein the second container and the third container are positioned in a second portion of the sample preparation system.

24. The sample preparation system of claim 23, wherein the first portion and the second portion are removably coupled to one another.

25. The sample preparation system of claim 23 or claim 24, wherein the second container and the third container are positioned in the second portion and are separated from one another by at least one of a seam and a weld in the second portion.

26. The sample preparation system of any one of claims 13-24, wherein the second container and the third container are separated by at least one of a seam and a weld.

27. The sample preparation system of any preceding claim, wherein the first container is adapted to be closed.

28. The sample preparation system of any preceding claim, wherein the first container comprises a bag.

29. The sample preparation system of any preceding claim, wherein the first container is freestanding.

30. The sample preparation system of any preceding claim, wherein at least one of the first container and the second container includes a port.

31. The sample preparation system of any preceding claim, wherein the second container is adapted to be removably coupled to the first container.

32. The sample preparation system of any preceding claim, wherein the second container is collapsible.

33. The sample preparation system of any preceding claim, wherein the second container is adapted for incubation.

34. The sample preparation system of any preceding claim, wherein the second container is adapted to be coupled to a detection system.

35. The sample preparation system of any preceding claim, wherein the second container is adapted to be in fluid communication with the detection system.

36. A method for preparing samples, the method comprising: providing a first container containing a source, the first container being collapsible; agitating the source in the first container to form a liquid composition comprising the source; and moving at least a portion of the liquid composition from the first container to a second container coupled to the first container, the second container adapted to enrich a microorganism of interest in the liquid composition to form an enriched liquid composition.

37. The method of claim 36, further comprising agitating at least one of the source and the liquid composition in the second container.

38. The method of any one of claims 36-37, wherein agitating includes paddle blending.

39. The method of any one of claims 36-38, wherein moving at least a portion of the liquid composition includes moving at least a portion of the liquid composition without exposing the liquid composition to ambience.

40. The method of claim 39, wherein moving at least a portion of the liquid composition without exposing the liquid composition to ambience includes actuating a valve.

41. The method of any one of claims 36-40, further comprising adding a diluent to the first container, and wherein agitating the source includes agitating the source and the diluent to form the liquid composition.

42. The method of any one of claims 36-41, further comprising filtering the liquid composition to form a filtrate, and wherein moving at least a portion of the liquid composition includes moving at least a portion of the filtrate to a second container.

43. The method of any one of claims 36-42, further comprising enriching the microorganism of interest in the second container to form the enriched liquid composition.

44. The method of any one of claims 36-43, further comprising testing for at least one of the presence, quantity and viability of the microorganism of interest.

45. The method of any one of claims 36-44, further comprising incubating the second container.

46. The method of any one of claims 36-45, further comprising removing at least a portion of the liquid composition from the second container.

47. The method of any one of claims 36-46, further comprising coupling the second container to a detection system.

48. The method of any one of claims 36-47, further comprising moving at least a portion of the liquid composition from at least one of the first container and the second container to a third container, the third container being coupled to at least one of the first container and the second container.

49. The method of claim 48, wherein moving at least a portion of the liquid composition to the third container includes moving at least a portion of the liquid composition to the third container without exposing the liquid composition to ambience.

50. The method of any one of claims 48-49, further comprising enriching the microorganism of interest in the third container.

51. The method of any one of claims 48-50, wherein the microorganism of interest is a first microorganism of interest, and further comprising enriching a second microorganism of interest in the third container, wherein the second microorganism of interest is different from the first microorganism of interest.

52. The method of any one of claims 48-51, wherein the first container is positioned in a first portion of the sample preparation system and the second container and the third container are positioned in a second portion of the sample preparation system, and further comprising decoupling the first portion and the second portion.

53. The method of any one of claims 36-52, further comprising coupling the second container to the first container.

54. The method of claim 53, wherein coupling the second container to the first container includes providing fluid communication between the first container and the second container.