WO2023236736A1 - Sample collection apparatus and method, and sample processing apparatus and method - Google Patents

Abstract

A sample collection apparatus, comprising: a mouthpiece (101, 301), wherein the mouthpiece (101, 301) is configured to engage with at least a portion of the mouth of a subject; a length-adjustable component (102, 302), wherein a first end of the length-adjustable component (102, 302) is coupled to the mouthpiece (101, 301), and the length-adjustable component (102, 302) has a first state and a second state, wherein in the first state, the length-adjustable component (102, 302) has a first length along the longitudinal axis thereof, and in the second state, the length-adjustable component (102, 302) has a second length along the longitudinal axis, with the second length greater than the first length; and a sampling head (103, 303), wherein the sampling head (103, 303) is coupled to a second end, substantially opposite to the first end along the longitudinal axis, of the length-adjustable component (102, 302), and the sampling head (103, 303) is configured, when the length-adjustable component (102, 302) is in the second state, to contact a collection site in the subject and collect a biological sample from the collection site.

Description

Sample collection device and method and sample processing device and method

cross reference

This application claims the benefit of International Application No. PCT/CN2022/097130, filed on June 6, 2022, which is incorporated herein by reference in its entirety.

Technical field

The present disclosure relates to devices and methods for sampling samples, as well as devices and methods for processing samples.

Background technique

A swab is usually used to sample a biological sample from the subject's oropharynx, and then the swab head with the subject's biological sample is broken off and the sampling head is placed in a reaction tube, and then the reaction tube is sampled. Processing of biological samples, such as nucleic acid amplification and detection. Nucleic acid amplification methods can selectively amplify and identify nucleic acids of interest from complex mixtures, such as biological samples. In order to detect nucleic acids in a biological sample, the biological sample is typically processed to separate the nucleic acid from other components of the biological sample and other substances that may interfere with the nucleic acid and/or amplification. After isolating the nucleic acid of interest from the biological sample, the nucleic acid of interest can be amplified by, for example, amplification methods known in the art, such as thermal cycle-based methods (eg, polymerase chain reaction (PCR)). increase. After the nucleic acid of interest is amplified, the amplification products can be detected and the detection results interpreted by the end user.

Contents of the invention

In one aspect, the invention provides a sample collection device. The sample collection device may include: a mouthpiece configured to engage at least a portion of a subject's mouth; a variable length member having a first end coupled to the mouthpiece and having a length of The variable length member has a first state and a second state. In the first state, the variable length member has a length along its longitudinal axis. a first length of wire, the variable length member having a second length along the longitudinal axis in the second state, the second length being greater than the first length; and a sampling head coupled to the variable length member along the longitudinal axis. At a second end substantially opposite the first end, the sampling head is configured to contact the collection site of the subject and collect a biological sample from the collection site when the variable length member is in the second state.

In another aspect, the invention provides a method of collecting a biological sample from a subject. The method may include: (a) providing a sample collection device including (i) a mouthpiece member, (ii) a variable length member, a first end of the variable length member coupled to the mouthpiece member, wherein the variable length member is in a first state having a first length along its longitudinal axis, and (iii) a sampling head coupled to a second end of the variable length member substantially opposite the first end along the longitudinal axis, wherein the mouthpiece engages at least a portion of the subject's mouth; (b) extending the variable length member with the mouthpiece engaged at least a portion of the subject's mouth such that the variable length member extends from the first The state changes to a second state, wherein in the second state the variable length member has a second length along the longitudinal axis, the second length being greater than the first length, wherein in the second state of the variable length member the sampling head is in contact with the receiver contact the subject's collection site to thereby collect the subject's biological sample from the collection site; and (c) remove the mouthpiece member from the subject's mouth.

In another aspect, the invention provides a kit. The kit may include: a sample collection device according to an aspect of the present disclosure; and optionally, instructions to guide a user to use the sample collection device to collect a biological sample from a subject.

In another aspect, the present invention provides a method of sample collection. The method may include: (a) providing a sample collection device comprising (i) a mouthpiece member configured to engage at least a portion of the subject's mouth, (ii) a variable length member , a first end of the variable length member is coupled to the mouthpiece member, the variable length member has a first state and a second state, in the first state, the variable length member has a first length along its longitudinal axis, and in In the second state, the variable length member has a second length along the longitudinal axis, the second length being greater than the first length, and (iii) a sampling head coupled to the first end of the variable length member along the longitudinal axis. a substantially opposite second end, a sampling head configured to contact a collection site of the subject and collect a biological sample from the collection site when the variable length member is in the second state, wherein the sampling head includes a biological sample of the subject; and (b) place the sampling head in the container.

In another aspect, the invention provides a sample collection device. The sample collection device may include: a mouthpiece configured to engage at least a portion of the subject's mouth; a swab support coupled to the mouthpiece and configured to support the swab, wherein the swab has a first position relative to the mouthpiece member and a first position different from the first position, wherein the swab when in the second position contacts a collection site of the subject and collects the biological sample from the collection site .

In another aspect, the invention provides a method of collecting a biological sample from a subject. The method may include: (a) providing a sample collection device including (i) a mouthpiece member, and (ii) a swab support member coupled to the mouthpiece member and configured to support the swab , wherein the swab has a first position relative to the mouthpiece member, wherein the mouthpiece member engages at least a portion of the subject's mouth; (b) where the mouthpiece member engages at least a portion of the subject's mouth changing the swab from a first position to a second position relative to the mouthpiece member, the second position being different from the first position, wherein the swab when in the second position is in contact with the subject's collection site and Collect a biological sample from the collection site; and (c) remove the mouthpiece member from the subject's mouth.

In another aspect, the present invention provides a sample processing device. The sample processing device may include: a container having an opening at one end, the container being configured to receive a biological sample therein; and a cover configured to seal the opening of the container and including receiving therein at least one component necessary for processing the biological sample. At least one chamber for reagents, the at least one chamber including a reagent release member that when activated releases at least one reagent into the container.

In another aspect, the invention provides a method of processing a biological sample. The method may include: (a) providing a sample processing device, the sample processing device comprising (i) a container having an opening at one end, the container being configured to receive a biological sample therein, and (ii) a closure configured to sealing the opening of the container and including at least one chamber containing at least one reagent necessary for processing the biological sample; (b) placing the biological sample within the container through the opening; (c) coupling the closure to the container opening; and (d) activating a reagent release member thereby releasing at least one reagent into the container.

Other aspects and advantages of the invention will become apparent to those skilled in the art from the following detailed description, in which only illustrative embodiments of the invention are shown and described. It will be appreciated that the present disclosure is capable of other and different embodiments, and Its several details are capable of modifications in various obvious respects, all without departing from the present disclosure. Accordingly, the drawings and descriptions are to be considered illustrative rather than restrictive.

Incorporation by reference

All publications, patents and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference.

Description of the drawings

The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the invention will be obtained by reference to the following detailed description, illustrating illustrative embodiments in which the principles of the invention are utilized, and the accompanying drawings, in which:

Figure 1 is a schematic diagram depicting an exemplary sample collection device of the present disclosure.

2A and 2B are schematic diagrams depicting first and second states, respectively, of an exemplary sample collection device of the present disclosure.

Figure 3 is a schematic diagram depicting yet another exemplary sample collection device of the present disclosure.

4 is a flowchart depicting an exemplary method of sampling a biological sample from a subject using a sample collection device of the present disclosure.

Figure 5 is a schematic diagram depicting a sample preservation and/or processing device consistent with the present disclosure.

Figure 6 is a flowchart depicting an exemplary method for sample collection of the present disclosure.

7A and 7B are schematic diagrams illustrating yet another exemplary sample collection device of the present disclosure.

Figure 8 is a cross-sectional view illustrating yet another exemplary sample collection device of the present disclosure.

Figure 9 is yet another schematic diagram illustrating yet another exemplary sample collection device of the present disclosure.

10 is a flowchart depicting an exemplary method of sampling a biological sample from a subject using yet another sample collection device of the present disclosure.

Figure 11 is a schematic diagram depicting a sample processing device of the present disclosure.

Figure 12 is yet another schematic diagram depicting a sample processing device of the present disclosure.

Figure 13 is a flow chart depicting an exemplary method of processing a biological sample of the present disclosure.

Figure 14 illustrates a computer system programmed or otherwise configured to implement a sample collection method or sample processing method provided by the present disclosure.

Detailed ways

While various embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Many variations, modifications and substitutions will occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed.

As used in this specification and the claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, the term "a cell" includes a plurality of cells, including mixtures thereof.

As used herein, the term "amplification" generally refers to the production of one or more copies of a nucleic acid or "amplification product." The term "DNA amplification" generally refers to the generation of one or more copies of a DNA molecule or an "amplified DNA product." The term "reverse transcription amplification" generally refers to the generation of deoxyribonucleic acid (DNA) from a ribonucleic acid (RNA) template through the action of reverse transcriptase.

As used herein, the term "denaturation" generally refers to the complete or partial unwinding of the helical structure of double-stranded nucleic acids, and in some cases to the unwinding of the secondary structure of single-stranded nucleic acids. Denaturation can include inactivation of the pathogen cell wall or viral coat, as well as inactivation of inhibitor proteins. The conditions under which denaturation can occur include "denaturation temperature" which generally refers to the temperature at which denaturation is allowed to occur, and "denaturation duration" which generally refers to the amount of time allotted for denaturation to occur.

As used herein, the term "extension" generally refers to the incorporation of nucleotides into nucleic acids in a template-directed manner. Extension can occur with the aid of enzymes such as, for example, polymerases or reverse transcriptases. The conditions under which extension can occur include "extension temperature" which generally refers to the temperature at which extension is allowed to occur, and "extension duration" which generally refers to the amount of time allotted for extension to occur.

As used herein, the term "nucleic acid" generally refers to a polymeric form of nucleotides (deoxyribonucleotides (dNTPs) or ribonucleotides (rNTPs)) or analogs thereof of any length. Nucleic acids can have any three-dimensional structure and can perform any known or unknown function. Non-limiting examples of nucleic acids include DNA, RNA, coding or non-coding regions of a gene or gene fragment, one or more loci determined by linkage analysis, exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, short interfering RNA (siRNA), short hairpin RNA (shRNA), microRNA (miRNA), ribozyme, cDNA, recombinant nucleic acid, branched nucleic acid, plasmid, vector, isolated any Sequenced DNA, isolated RNA of any sequence, nucleic acid probes and primers. Nucleic acids may contain one or more modified nucleotides, such as methylated nucleotides and nucleotide analogs. Modifications to the nucleotide structure, if present, can be performed before or after nucleic acid assembly. The nucleotide sequence of a nucleic acid can be interrupted by non-nucleotide components. The nucleic acid can be further modified after polymerization, for example by coupling or conjugation with a reporter agent.

As used herein, the term "primer extension reaction" generally refers to the denaturation of a double-stranded nucleic acid, the binding of a primer to one or both strands of the denatured nucleic acid, and subsequent extension of the primer.

As used herein, the term "reaction mixture" generally refers to a composition that includes reagents necessary to accomplish nucleic acid amplification (e.g., DNA amplification, RNA amplification), non-limiting examples of such reagents include target RNA Or the target DNA has a specific primer set, DNA produced by reverse transcription of RNA, a DNA polymerase, a reverse transcriptase (for example, for reverse transcription of RNA), a suitable buffer (including a zwitterionic buffer), an auxiliary factors (e.g., divalent and monovalent cations), dNTPs, and other enzymes (e.g., uracil-DNA glycosylase (UNG), etc.). In some cases, the reaction mixture may also contain one or more reporters.

As used herein, the term "target nucleic acid" generally refers to a nucleic acid molecule having a certain nucleotide sequence in a starting population of nucleic acid molecules, its presence, amount and/or sequence, or one or more of these Changes need to be measured. The target nucleic acid can be any type of nucleic acid, including DNA, RNA, and their analogs. As used herein, "target ribonucleic acid (RNA)" generally refers to a target nucleic acid that is RNA. As used herein, "target deoxyribonucleic acid (DNA)" generally refers to a target nucleic acid that is DNA.

As used herein, the term "subject" generally refers to an entity or medium having testable or detectable genetic information. The subject may be a person or an individual. The subject may be a vertebrate animal, such as a mammal. Non-limiting examples of mammals include rats, monkeys, humans, livestock, athletic animals, and pets. Other examples of subjects include food, plants, soil, and water.

As used herein, the term "biological sample" generally refers to any sample taken from a target location in a subject. Non-limiting examples of biological samples include blood (or components of blood—e.g., White blood cells, red blood cells, platelets), cells obtained from any anatomical location of the subject, skin, heart, lungs, kidneys, exhaled breath, bone marrow, feces, semen, vaginal fluid, interstitial fluid derived from tumor tissue, breast, pancreas , cerebrospinal fluid, tissue, throat swab, biopsy, placental fluid, amniotic fluid, liver, muscle, smooth muscle, bladder, gallbladder, colon, intestine, brain, luminal fluid, sputum, pus, microbiota, meconium, breast milk , prostate, esophagus, thyroid, serum, saliva, urine, gastric and digestive juices, tears, eye fluids, sweat, mucus, earwax, oil, glandular secretions, spinal fluid, hair, nails, skin cells, plasma, Nasal swabs or nasopharyngeal washes, spinal fluid, cord blood, and/or other excreta or body tissue.

As used herein, the term "oropharynx", also known as the middle pharynx or oropharynx, generally refers to the portion of the subject located between the free edge of the velum palatini and the plane of the superior edge of the epiglottis, which communicates anteriorly with the oral cavity via the pharyngeal isthmus. , continues the nasopharynx superiorly, and connects the laryngopharynx inferiorly.

Figure 1 is a schematic diagram depicting an exemplary sample collection device of the present disclosure. 2A and 2B are schematic diagrams depicting first and second states, respectively, of an exemplary sample collection device of the present disclosure. The sample collection device may include a mouthpiece member 101 , a variable length member 102 coupled to the mouthpiece member 101 at a first end of its longitudinal axis, and a variable length member 102 coupled to the variable length member 102 along the longitudinal axis substantially dissimilar to the first end. Opposite the second end of the sampling head 103 . The sample collection device is shaped and sized for placement in a subject's oral cavity.

The mouthpiece member 101 of the sample collection device is configured to engage at least a portion of the subject's mouth. During use of the exemplary sample collection device, the mouthpiece is positioned outside at least a portion of the subject's mouth. In some cases, the at least a portion of the mouth includes the subject's lips. The mouthpiece member is shaped and sized to contact the subject's lips outside the subject's oral cavity, thereby preventing the subject from accidentally swallowing the sample collection device. The shape of the mouthpiece piece may be circular, oval, rectangular, or square. In one example, the mouthpiece piece may be a rectangular shaped piece with chamfers. The mouthpiece member may have one or more openings to ensure that the external air environment communicates with the subject's respiratory system via the openings during use of the sample collection device. In one example, one or more openings may be disposed near the geometric center of the mouthpiece member.

In some cases, the mouthpiece piece may be made of a resilient material such as silicone. In other cases, the mouthpiece piece may be made from a substantially rigid material, such as plastic. In other cases, the mouthpiece member may include laminated layers of material, such as a layer of elastic material and a layer of substantially rigid material. In one example, the mouthpiece piece can have a layer of elastic material proximally relative to the subject and a substantially rigid layer of material distally relative to the subject. In one example, the mouthpiece member may have a handle distally relative to the subject to facilitate a user (eg, subject, healthcare provider) holding the sample collection device.

The variable length component 102 of the sample collection device of the present disclosure has a first end and a second end along its longitudinal axis. The variable length member has a first end (ie, distal end relative to the subject) coupled to the mouthpiece 101 and a second end (ie, proximal end relative to the subject) coupled to the sampling head 103 . The length of the variable length component is variable along its longitudinal axis. For example, the variable length component may have a first state (eg, as shown in Figure 2A) and a second state (eg, as shown in Figure 2B). In a first state, the variable length member has a first length along its longitudinal axis; and in a second state, the variable length member has a second length along said longitudinal axis. The second length may be greater than the first length. In some cases, the first state may be a state in which no external force exceeding a preset threshold is applied to the variable-length component, and the second state may be a state in which an external force exceeding a preset threshold is applied to the variable-length component. state.

In some cases, the variable length component 102 is deformable when a force exceeding a predetermined threshold is exerted on the variable length component 102 . This deformation of the variable length component results in a change in its length along the longitudinal axis. In the example shown in Figures 1, 2A and 2B, the variable length member includes two or more deformable ribs extending substantially along its longitudinal axis. The deformable ribs enclose a space in their middle. The deformable ribs may be made of elastic material. In the first state of the variable length component, at least a portion of the deformable rib may be curved along the longitudinal axis of the variable length component. When an external force exceeding a preset threshold is applied to the deformable rib, the deformable rib deforms, so that the length of the deformable rib along the longitudinal axis of the variable-length component changes, as shown in FIG. 2B . The external force may be applied in a direction substantially perpendicular to the longitudinal axis of the variable length component such that deformation of the variable length component causes the deformable rib to move inwardly. For example, in one example, when using the sample collection device of the present disclosure, the variable length member is positioned within the subject's mouth; the subject then closes the mouth so that at least the subject's teeth, upper jaw, lower jaw, inner The cheek or combination thereof exerts an external force on the variable-length component, causing the variable-length component to The deformable ribs deform and change in length. In some cases, the deformable rib is deformable at least in a portion proximate the mouthpiece member such that the deformable rib occurs when the subject closes the mouth such that an external force is exerted on the deformable rib by at least the subject's teeth. deforms and causes its length to change. In other cases, the deformable rib is deformable at least midway along its length such that when the subject closes the mouth such that an external force is exerted on the deformable rib by at least the subject's upper and lower jaws, the deformable rib may The deformation rib deforms and causes its length to change.

The length change value of the variable length component of the sample collection device of the present disclosure may be a preset value or within a preset range. When the subject applies force to the variable length component by closing the mouth, the length change value of the variable length component along the longitudinal axis is preset or within a preset range. The preset value can be at least 0.1 centimeters (cm), 0.2cm, 0.4cm, 0.6cm, 0.8cm, 1.0cm, 1.2cm, 1.4cm, 1.6cm, 1.8cm, 2.0cm, 2.2cm, 2.4cm, 2.6 cm, 2.8cm, 3.0cm, 3.2cm, 3.4cm, 3.6cm, 3.8cm, 4.0cm, 4.2cm, 4.4cm, 4.6cm, 4.8cm, 5.0cm, 5.2cm, 5.4cm, 5.6cm, 5.8cm, 6.0cm, 6.2cm, 6.4cm, 6.6cm, 6.8cm, 7.0cm, 7.2cm, 7.4cm, 7.6cm, 7.8cm, 8.0cm, 8.2cm, 8.4cm, 8.6cm, 8.8cm, 9.0cm, 9.2cm , 9.4cm, 9.6cm, 9.8cm, 10.0cm, 10.5cm, 11.0cm, 11.5cm, 12.0cm, 12.5cm, 13.0cm, 13.5cm, 14.0cm, 14.5cm or 15.0cm. This preset value can be greater than any of the above preset values. The preset value can be a value between any two preset values mentioned above. The preset range may be a range between any two preset values mentioned above. The length change value or transformation range of the length-variable component can be achieved by setting the shape and/or size of the length-variable component. For example, the length variation value or variation range of the variable length component can be controlled by controlling the initial bending degree and initial length of the deformable rib of the variable length component. Various models of sample collection devices can be provided, with variable length components having different length change values or conversion ranges, so that they can be used by subjects of different ages, genders, and oral cavity sizes.

The direction in which the length of the length-variable component of the sample collection device of the present disclosure changes can be preset. For example, when a subject applies force to the variable length component by closing the mouth, the change in length of the variable length component may occur in a preset direction. In some cases, the change in length of the variable length component may occur substantially along its longitudinal axis. In other cases, the length of the variable length component may occur in a direction offset from its longitudinal axis. The direction in which the length of the variable-length component changes can be achieved by controlling the shape and material of the variable-length component. For example, the direction in which the length of the variable length component changes may be controlled by controlling the initial degree of curvature, initial length, material, or a combination thereof of each of the plurality of deformable ribs of the variable length component. In one example, the initial lengths of the plurality of deformable ribs of the variable length component may be different. When the subject applies force to the variable-length component by closing the mouth, the deformable ribs with the smaller initial lengths undergo smaller changes in length, causing the length changes of the variable-length component to occur in the longitudinal direction away from the variable-length component. in the direction of the axis. In another example, the plurality of deformable ribs of the variable length component may be made of different materials. Deformable ribs made of materials with smaller deformation properties undergo smaller changes in length when a subject applies force to the variable-length component by closing the mouth, causing the length change of the variable-length component to occur at an offset length in the direction of the longitudinal axis of the variable component.

The sampling head 103 of the sample collection device of the present disclosure is coupled to the second end (ie, the proximal end relative to the subject) of the variable length member 102 . The sampling head is configured to contact a collection site of a subject and collect a biological sample from the collection site when the variable length member is in the second state having an elongated longitudinal length. When the length-variable component is in the second state, the sampling head exerts a force above a preset threshold on the subject's collection site, thereby ensuring that the sampling head effectively collects biological samples from the subject's collection site. In some examples, the sampling head can be removed from the end of the variable length component. The sampling head may include materials suitable for transferring a biological sample (eg, saliva, oropharyngeal secretions, nasopharyngeal secretions) to and retained by the sampling head. Exemplary materials may include, but are not limited to, cotton, polyester, polyester, rayon, and the like. In one example, these materials can be formed into a material layer fixed on the end of the sampling head using processes such as winding, spraying, and flocking. In another example, these materials may be formed into bristles affixed to the end of the sampling head.

Figure 3 is a schematic diagram depicting yet another exemplary sample collection device of the present disclosure. The sample sampling device of this embodiment may include a mouthpiece 301, a variable length member 302 coupled to the mouthpiece 301 at a first end of its longitudinal axis, a variable length member 302 coupled to the variable length member 302 along the longitudinal axis and a third end thereof. One end is substantially opposite to the second end of the sampling head 303. The mouthpiece member 301 , variable length member 302 and sampling head 303 of the sample collection device of this embodiment may be the same as the mouthpiece member, variable length member of the sample collection device of the embodiment described with reference to FIGS. 1 , 2A and 2B Hecai The structure of the sample heads is basically the same. The variable length component 302 of the sample collection device of this embodiment may further include a rod 3021 extending substantially along the longitudinal axis of the variable length component. In one example, rod 3021 may extend between two or more deformable ribs and the sampling head.

At least a portion of the rod may be made of elastic material. In some cases, when a force exceeding a threshold is applied to the rod, at least a portion of the rod is deformable to conform to the intraoral structure of the subject and increase the subject's comfort during the sampling process. The bending direction of the rod is preset when a force exceeding a threshold is applied to the rod. When a force exceeding a threshold is applied to the rod, the degree of bending of the rod is preset or within a preset range. This controlled bending of the rod can be achieved by adjusting the material the rod is made of. For example, different circumferential portions of the rod may be made of materials that allow different amounts of deformation (e.g., materials that stretch differently along the longitudinal axis of the rod), such that when an external force is applied to the rod, the rod only moves in the direction allowed by the rod. Parts made of materials with low deformation are bent.

The length of the rod may be at least 0.1cm, 0.2cm, 0.4cm, 0.6cm, 0.8cm, 1.0cm, 1.2cm, 1.4cm, 1.6cm, 1.8cm, 2.0cm, 2.2cm, 2.4cm, 2.6cm, 2.8cm , 3.0cm, 3.2cm, 3.4cm, 3.6cm, 3.8cm, 4.0cm, 4.2cm, 4.4cm, 4.6cm, 4.8cm, 5.0cm, 5.2cm, 5.4cm, 5.6cm, 5.8cm, 6.0cm, 6.2 cm, 6.4cm, 6.6cm, 6.8cm, 7.0cm, 7.2cm, 7.4cm, 7.6cm, 7.8cm, 8.0cm, 8.2cm, 8.4cm, 8.6cm, 8.8cm, 9.0cm, 9.2cm, 9.4cm, 9.6cm, 9.8cm, 10.0cm, 10.5cm, 11.0cm, 11.5cm, 12.0cm, 12.5cm, 13.0cm, 13.5cm, 14.0cm, 14.5cm or 15.0cm. The length of the rod can be greater than any of the above values. The length of the rod can be a value between any two of the above values. Several models of sample collection devices are available with variable-length shafts of varying lengths for use with subjects of different ages, genders, and oral cavity sizes.

Figure 4 is a flowchart depicting an exemplary method of collecting a biological sample from a subject of the present disclosure. In process 401, a sample collection device is provided. In process 402, the sampling head of the sample collection device is caused to collect the biological sample from the collection site of the subject. In process 403, the sample collection device is removed from the subject's mouth.

In process 401, the provided sample collection device may be the sample collection device of the exemplary embodiment described above with reference to FIGS. 1, 2A and 2B or the sample collection device described with reference to FIG. 3. Sample collection device of an exemplary embodiment. In one example, a sample collection device may include a mouthpiece member, a variable length member, and a sampling head. In step 401, optionally, a sample collection device may be placed into the subject's oral cavity. At this point, the variable length member is in a first, unstretched state having a first length along its longitudinal axis. The mouthpiece engages at least a portion of the subject's mouth (eg, lips).

In process 402, with the mouthpiece engaged with at least a portion of the subject's mouth, a variable length component of the sample sampling device is extended such that the variable length component changes from a first state (i.e., an unstretched state ) changes to the second state (i.e., elongated state). In the second state, the variable length member has a second length along the longitudinal axis that is greater than the first length. In the second state of the variable length component, the sampling head of the sample sampling device is in contact with the collection site of the subject to collect the subject's biological sample (eg, saliva, oropharyngeal secretions containing nucleic acids) from the collection site. wait). In this process, for example, the subject closes the mouth such that a force is exerted on the variable length component of the sample sampling device by at least the subject's teeth, upper jaw, lower jaw, inner cheek, or a combination thereof, resulting in the variable length component Length changes occur. This change in length of the variable length member allows the sampling head located at the end of the variable length member to reach the subject's site. In one example, the subject's collection site may be the oropharyngeal site. When using the sample collection device of the present disclosure to collect a biological sample at the oropharyngeal site of a subject, the subject puts the sample collection device into the mouth, and then the subject applies force to the variable length member by closing the mouth , causing the length of the variable-length component to change in the direction of its longitudinal axis, thereby allowing the sampling head to reach the oropharynx of the subject. During the change of the variable length member from the first state to the second state and during the time the variable length member remains in the second state, at least a portion of the proximal side of the mouthpiece member abuts the subject's lip, thereby preventing There is a danger if the subject swallows the entire sample collection device in his mouth.

In some cases, process 402 may be performed more than once, such that the sampling head contacts the subject's collection site multiple times to ensure that an effective amount of biological sample is collected from the subject's collection site. In some cases, the variable length component may be maintained in the second state for a preset time. For example, the subject keeps the mouth closed and the sampling head continues to contact the subject's collection site for a preset time, thereby ensuring that an effective amount of biological sample is collected from the subject's collection site. The preset time may be at least 0.5 seconds (sec), 1.0sec, 2.0sec, 3.0sec, 4.0sec, 5.0sec, 6.0sec, 7.0sec, 8.0sec, 9.0sec, 10.0sec, 11.0sec, 12.0sec, 13.0sec, 14.0sec, 15.0sec, 16.0sec, 17.0sec, 18.0sec, 19.0sec, 20.0sec, 21.0sec, 22.0sec , 23.0sec, 24.0sec, 25.0sec, 26.0sec, 27.0sec, 28.0sec, 29.0sec, 30.0sec, 35.0sec, 40.0sec, 45.0sec, 50.0sec, 55.0sec, 1 minute (min), 2min, 3min, 4min or 5min. The preset time can be greater than any of the above values. The preset time can be a value between any two values mentioned above. The preset time may vary according to factors such as the collection site to be sampled, the biological sample to be collected, the processing of the biological sample to be performed, the inspection of the biological sample to be performed, the parameters and requirements of the inspection, and other factors.

After the collection of the biological sample is completed, the sample collection device is removed from the subject's oral cavity in process 403 . For example, the subject or healthcare provider may hold the portion of the mouthpiece outside the subject's lips (e.g., the perimeter of the flap portion of the mouthpiece, or a brow grip distal to the mouthpiece) , remove the sample collection device from the subject's mouth.

The present disclosure also provides a kit, which includes the sample collection device of the present disclosure and instructions for guiding a user to use the sample collection device to collect a biological sample of a subject.

Figure 5 is a schematic diagram depicting a sample preservation and/or processing device consistent with the present disclosure. The sample retention and/or processing device may include a container 501 and a closure 502 . Container 501 has an opening at one end. Lid 502 is configured to couple to and seal the opening of container 501 . The closure may be, for example, a screw cap, a snap cap or a flip cap.

In some examples, the container of the sample storage and/or processing device of the present disclosure may be pre-filled with sample processing fluid. The sample processing solution and the biological sample are mixed in proportion to obtain the processed sample. In some examples, a container of a sample preservation and/or processing device of the present disclosure may be configured to have at least one reagent pre-stored therein necessary for preservation and/or processing of a biological sample. In some examples, at least one reagent includes all reagents necessary to complete sample preservation, sample processing, nucleic acid amplification (eg, DNA amplification, RNA amplification). Reagents necessary for processing biological samples include those necessary for reverse transcription and nucleic acid amplification (e.g., reverse transcriptase, DNA polymerase, dNTPs, cofactors, primers, appropriate buffers, etc.) and reporter reagents (e.g., containing FAM Dye oligonucleotide probes). After adding the biological sample collected from the subject, all processing of the sample required for nucleic acid amplification can be completed in the container.

In some embodiments, the biological sample is stored and/or processed as provided in the present disclosure. not yet purified in the processing unit. In some embodiments, when the biological sample is provided to the sample preservation and/or processing device of the present disclosure, the nucleic acid of the biological sample has not yet been extracted. For example, when a biological sample is provided to a sample preservation and/or processing device of the present disclosure, RNA or DNA in the biological sample may not be extracted from the biological sample. Additionally, in some embodiments, target nucleic acids (eg, target RNA or target DNA) present in the biological sample may not be concentrated prior to providing the biological sample to a sample preservation and/or processing device of the present disclosure.

The sample preservation and/or processing devices of the present disclosure may be used in conjunction with the sample collection devices of the present disclosure. In some examples, the cover may further include a sample holder 503 configured to retain at least a portion of the sampling head 103 of the sample collection device of the present disclosure. For example, the sampling head 103 of the sample collection device of the present disclosure may be removed from the end of the variable length member. Correspondingly, the inner side of the cover of the sample storage and/or processing device of the present disclosure (ie, with the opening of the container of the sample storage and/or processing device) is provided with a sampling head capable of fixing the sample collection device of the present disclosure (which e.g. The sample holder may be at least a portion of a sphere. The sample holder may be a recess, a clamp, or the like. For example, the sample holder may include a plurality of resilient arms defining a space therebetween for receiving at least a portion of the sampling head. In one example, after completing collection of a biological sample from a subject's collection site, a user (eg, the subject himself, or the subject's medical care provider) may hold the mouthpiece member of the sample collection device , inserting the sampling head of the sample collection device into the capped sample holder of the sample preservation and/or processing device of the present disclosure, thereby transferring and securing the removable sampling head carrying the biological sample of the subject to the present disclosure. within the cover of the sample storage and/or processing device.

In some examples, the capped sample holder is configured to transfer at least a portion of a collected biological sample from a sampling head of a sample collection device of the present disclosure to the sample holder. For example, the capped sample holder includes a hygroscopic material that adsorbs to and retains therein at least a portion of the collected biological sample from the sampling head of the sample collection device of the present disclosure. In one example, after completing collection of a biological sample from a subject's collection site, a user (eg, the subject himself, or the subject's medical care provider) may hold the mouthpiece member of the sample collection device , placing the sampling head of the sample collection device against the capped sample holder of the sample preservation and/or processing device of the present disclosure. the water-absorbent material of the sample holder adsorbs from the sampling head at least a portion of the biological sample of the subject carried by the sampling head, Transfer of the biological sample from the sampling head to the cover of the sample preservation and/or processing device of the present disclosure is thereby achieved.

After completing the transfer of the biological sample from the sample collection device of the present disclosure to the closure of the sample retention and/or processing device, the user couples the closure to the opening of the container of the sample retention and/or processing device, thereby sealing the opening . The user then only needs to shake the sample storage and/or processing device so that the reagents in the container come into contact with the removable sampling head or sample holder in the cover, and the removable sampling head or sample holder can be retained on the cap. The subject's biological samples are rinsed and transferred to preset reagents in the sample storage and/or processing device.

In some cases, a filter may be provided at the opening of or within the container of the sample storage and/or processing device of the present disclosure, and the area of the filter may be substantially the same as the cross-section of the container. The filter is configured to filter out impurities from the biological sample. For example, where the sampling head is fabricated by flocking, a portion of the fibrous material on the sampling head may become detached during transfer of the sampling head to the cover of the sample holding and/or processing device and be transported along with the biological material. The samples are flushed together into preset reagents in the sample storage and/or processing device. The filter prevents fibrous tissue from entering subsequent sample processing stages. The pore size of the filter mesh may vary depending on the type of biological sample collected, the material of the sampling head of the sample collection device, the processing to be performed on the sample, and so on.

Figure 6 is a flowchart depicting an exemplary method for sample collection of the present disclosure. In process 601, a sample collection device is provided. In process 602, the sampling head of the sample collection device is docked with a container.

In process 601, the provided sample collection device may be the sample collection device of the exemplary embodiment described above with reference to FIGS. 1, 2A and 2B or the sample collection device of the exemplary embodiment described with reference to FIG. 3. In one example, a sample collection device may include a mouthpiece member, a variable length member, and a sampling head. The mouthpiece may be configured to engage at least a portion of a subject's mouth. The first end of the variable length member may be coupled to the mouthpiece member. The variable length component may have a first state and a second state. In the first state, the variable length member may have a first length along its longitudinal axis. In the second state, the variable length member has a second length along the longitudinal axis. The second length may be greater than the first length. The sampling head may be coupled to a second end of the variable length member substantially opposite the first end along the longitudinal axis. The sampling head can be equipped with The variable length member is configured to contact the collection site of the subject and collect a biological sample from the collection site when the variable length member is in the second state. In step 401, a sample collection device may be placed into the subject's oral cavity. The subject then closes the mouth such that a force is exerted on the variable length component of the sample sampling device by at least the subject's teeth, maxilla, mandible, inner cheek, or a combination thereof, causing the variable length component to change in length. This change in length of the variable length member allows the sampling head located at the end of the variable length member to reach the subject's site. Finally, the sampling head collects and holds the subject's biological sample.

In process 602, the container used may be a sample holding and/or processing device as described with reference to Figure 5. The cover of the sample retention and/or processing device may further include a sample holder configured to retain at least a portion of the sampling head of the sample collection device of the present disclosure, or adapted to remove all of the sampling head from the sample collection device of the present disclosure. At least a portion of the collected biological sample is transferred to the sample holder. In one example, after completing collection of a biological sample from a subject's collection site, a user (eg, the subject himself, or the subject's medical care provider) may hold the mouthpiece member of the sample collection device , inserting the sampling head of the sample collection device into the capped sample holder of the sample preservation and/or processing device of the present disclosure, thereby transferring and securing the removable sampling head carrying the biological sample of the subject to the present disclosure. within the cover of the sample storage and/or processing device. The user then simply seals the cap to the opening of the container of the sample storage and/or processing device and shakes the sample storage and/or processing device to flush transfer the subject's biological sample to the sample storage and/or processing device. Pre-prepared reagents (for example, buffer) in the processing device.

7A and 7B are schematic diagrams illustrating yet another exemplary sample collection device of the present disclosure. FIG. 8 is a cross-sectional view showing the sample collection device. The sample collection device may include a mouthpiece member 701 and a swab support 702 coupled to the mouthpiece member 701 . The mouthpiece member 701 of the sample collection device is configured to engage at least a portion of the subject's mouth. During use of the sample collection device, at least a portion of the mouthpiece is located outside at least a portion of the subject's mouth. In some cases, the at least a portion of the mouth includes the subject's lips. The mouthpiece member is shaped and sized such that at least a portion thereof abuts the subject's lips outside the subject's oral cavity, thereby preventing the subject from accidentally swallowing the sample collection device. In the exemplary embodiment shown, the mouthpiece member may include a substantially cylindrical portion and an annular portion protruding from the cylindrical portion. During sample collection During use of the device, the annular portion of the mouthpiece abuts the subject's lips outside the subject's oral cavity, while the cylindrical portion is located in the subject's oral cavity and is clamped on the subject's upper and lower gums or upper and lower gums. between the teeth so that the mouthpiece piece is fixed relative to the subject's mouth. The mouthpiece piece may be made of elastic material such as silicone. When the subject bites the mouthpiece piece, the mouthpiece piece may deform to a certain extent.

The swab support 702 of the sample collection device is coupled to the mouthpiece 701 . In the embodiment shown in Figure 8, at least a portion of the swab support may be located within the interior space of the mouthpiece member and supported by the mouthpiece member. In one example, the swab support may include a cylindrical support portion with at least a portion of the cylindrical support extending through the interior space of the mouthpiece member along a longitudinal axis of the mouthpiece member and coupled with the mouthpiece member. The coupling of the swab support member and the mouthpiece member can be achieved in various ways. For example, one or more O-rings may be provided, which are sleeved on the outer periphery of the cylindrical support part perpendicularly to the longitudinal axis of the cylindrical support part of the swab support and are in contact with the inner wall or end of the mouthpiece member. part abuts, thereby fixing at least a part of the cylindrical support part of the swab support part in the internal space of the mouthpiece part. For another example, the swab support member can be clamped and fixed in the internal space of the mouthpiece member by a clamping structure (eg, clamping ring, clamping rib) provided inside the mouthpiece member. In another example, the swab support may include one or more O-rings that abut the interior or end of the mouthpiece member perpendicularly to the longitudinal axis of the mouthpiece member. The swab can be passed directly through the central opening of the O-ring, thereby being clamped and held by the O-ring. The orientation and/or position of the swab support relative to the mouthpiece may be changed. For example, the swab support may be secured in the interior space of the mouthpiece member by elastic clamping structures. When an external force is applied to the swab support, the orientation and/or position of the swab support relative to the mouthpiece member may be changed. For example, when an external force is applied to the swab support toward the subject, the swab support may move toward the subject relative to the mouthpiece member, and the swab support may also change orientation relative to the mouthpiece member.

The swab support 702 of the sample collection device is configured to support the swab 703 . In one example, the swab may axially pass through the cylindrical support portion of the swab support to be supported by the swab support. In another example, the swab can pass through the central opening of the o-ring of the swab support to be supported by the swab support. Swab 703 includes a swab head and a swab shaft coupled to the swab head. The swab head may include a material suitable for transferring a biological sample (eg, saliva, oropharyngeal secretions, nasopharyngeal secretions) to and retained by the swab head. Exemplary materials may include, but Not limited to, cotton, polyester, polyester or rayon, etc. A break point (eg, a cutout) may be provided on the swab shaft to facilitate the transfer of the swab head carrying the subject's biological sample to a sample storage device or sample processing device after sampling is completed.

In some embodiments, the swab can be displaced relative to the swab support such that the relative position of the swab relative to the swab support changes. For example, a user of the sample collection device (e.g., the subject or the subject's healthcare provider) may hold the distal end of the swab shaft of the swab and apply a force toward the subject, thereby causing the swab head to move relative to the subject. The swab support moves toward the subject's collection site. During this process, the position of the mouthpiece member and swab support member of the sample collection device relative to the subject's mouth may remain substantially unchanged. During this process, movement of the swab support relative to the mouthpiece may also occur, as described above. In some embodiments, the swab can have a first position and a second position relative to the swab support. When the swab is in the first position, the swab (eg, swab head) is not in contact with the subject's collection site. When the swab is in the second position, the swab (eg, swab head) contacts the collection site of the subject and collects a biological sample from the collection site. The subject's collection site may include the oropharyngeal site. The transition from the first position to the second position of the swab can be achieved by a user of the sample collection device operating the distal end of the swab shaft of the swab, or by an automated sampling device operating the swab shaft of the swab.

In some embodiments, the swab may be coupled to the swab support via an elastic member. For example, as shown in FIG. 8 , an elastic member 705 (for example, a spring) may be provided between the swab and the swab support. The elastic member is configured to control the onset and/or extent of displacement between the swab and the swab support. For example, the displacement of the swab relative to the swab support can only occur when the external force applied to the swab is large enough to deform the spring. When the external force applied to the swab reaches a preset threshold, the spring no longer deforms, causing the displacement of the swab relative to the swab support to stop, thereby preventing excessive displacement of the swab head from causing damage to the subject's collection site. damage. When the external force exerted on the swab disappears, the swab will return to its original position relative to the swab support. The elastic member may also be configured to provide feedback to the user such that the user tactilely senses the extent of displacement of the swab relative to the swab support.

In some embodiments, the swab can be coupled to the swab support through a buffer. For example, as shown in Figure 8, between the distal end of the swab (i.e., the end far away from the subject) and the distal end of the swab support Buffers 706 can be provided between the ends. The buffer member can be made of elastic material, or the buffer member can also be a spring. The buffer is configured to regulate the force exerted by the swab on the subject's collection site. For example, when the swab is in the second position, the swab head contacts the subject's collection site and exerts a certain degree of force on the subject's collection site. At this time, the swab rod can exert a reaction force on the buffering member in a direction away from the subject, thereby causing the buffering member to deform (eg, compress). Through the buffering effect brought by the deformation of the buffer member, the pressure value exerted by the swab head on the subject's collection site can be controlled within a preset range. In some embodiments, the sample collection device of the present disclosure may further include a handle coupled to the distal end of the swab (i.e., the end opposite the subject's collection site) to facilitate the user of the sample collection device to swab. sub to operate. The handle and the swab head of the swab may be respectively located at both ends of the mouthpiece member along the longitudinal axis of the mouthpiece member. In one example, the buffer member may be disposed between the distal end of the swab shaft and the proximal end of the handle.

In some embodiments, the sample collection device may also include a cover. As shown in FIG. 9 , the sample collection device may include a first cover 711 that may be configured to be removably coupled to a proximal end of the mouthpiece member along its longitudinal axis (i.e., proximal to the subject's one end) and cover at least the swab head portion of the swab, thereby ensuring that the swab head is not contaminated before using the sample collection device of the present disclosure. In some examples, the sample collection device may further include a second cover 712 that may be configured to be removably coupled to a distal end of the mouthpiece member along its longitudinal axis (i.e., distal to the subject's one end) and cover at least the handle. The removable coupling of the first cover and the second cover with the mouthpiece member may be achieved by snap-fitting (eg, protrusion/groove fitting), threaded fitting, or the like. In some examples, the sample collection device can be placed in a sterile disposable package. The user can unpack the swab and operate the swab by operating the swab support (eg, handle) to achieve sampling from the collection site of the subject. In some examples, the sample collection device is provided in the form of a kit. The kit may include a sample collection device as described above, and optionally instructions to guide the user to use the sample collection device to collect a biological sample.

10 is a flowchart depicting an exemplary method of sampling a biological sample from a subject using yet another sample collection device of the present disclosure. In process 1001, a sample collection device is provided. In process 1002, a biological sample is collected from a collection site of the subject using a swab of the sample collection device. In process 1003, the sample collection device is removed from the subject's mouth.

In process 1001, the sample collection device provided may be the exemplary sample collection device described above with reference to Figures 7A-9. In one example, the sample collection device may include a mouthpiece member and a swab support member. The swab support is coupled to the mouthpiece and configured to support the swab in a first position relative to the swab support. When the swab is in the first position, the swab head is not in contact with the subject's collection site.

In process 1002, the swab is changed from a first position to a second position relative to the swab support with the mouthpiece engaged with at least a portion of the subject's mouth (eg, lips), the The second position is different from the first position. When the swab is in the second position, the swab head contacts the subject's collection site and collects a biological sample from the collection site. In some cases, changing the swab from the first position to the second position may include causing a change in the relative position of the swab relative to the swab support. For example, a user of the sample collection device (eg, the subject or the subject's healthcare provider) may hold the distal end of the swab shaft of the swab and displace the swab head relative to the swab support. During this process, the positions of the sample collection device mouthpiece and the swab support relative to the subject's mouth are substantially unchanged, or displacement of the swab support relative to the mouthpiece may also occur. In some cases, changing the swab from the first position to the second position may also include changing the orientation of the swab support relative to the mouthpiece member. For example, the swab support is coupled to the mouthpiece through a deformable component (eg, an O-ring) such that when a user holds the swab with the distal end of the swab shaft substantially perpendicular to the swab support or When a force is exerted on the swab shaft and swab support in the direction of the longitudinal axis of the swab shaft, the force may cause a change in the orientation of the swab support relative to the mouthpiece member. The user can adjust the specific position of the swab head in contact with the collection site through this change in the orientation of the swab support member relative to the mouthpiece member, thereby achieving more accurate sampling.

In some examples, an elastic member (eg, a spring) disposed between the swab and the swab support may provide control and feedback of the onset and/or extent of displacement between the swab and the swab support. For example, the user is allowed to tactilely perceive the degree of displacement of the swab relative to the swab support. In some examples, a buffer disposed between the swab and the swab support may provide regulation of the force exerted by the swab on the subject's collection site. For example, when the swab head comes into contact with the collection site of the subject and exerts force on the collection site, the swab rod can apply a reaction force to the buffering member, thereby causing the buffering member to deform. Through the buffering effect brought by the deformation of the buffering parts, The pressure value exerted by the swab head on the subject's collection site can be controlled within a preset range.

In some cases, process 1002 may be performed more than once such that the swab head contacts the subject's collection site multiple times to ensure that an effective amount of biological sample is collected from the subject's collection site. In some cases, the swab can be held in the second position for a preset time.

After the collection of the biological sample is completed, the sample collection device is removed from the subject's oral cavity in process 1003 .

11 and 12 are schematic diagrams depicting sample processing devices of the present disclosure. The sample processing device may include a container 1101 and a processor cover 1102. Container 1101 has an opening at one end. Processor cover 1102 is configured to couple to and seal the opening of container 1101 . The processor cover may be, for example, a twist-on, snap-on or flip-up cover. Figure 11 shows a state where the processor cover is separated from the container, and Figure 12 shows a state where the processor cover is coupled to the container.

Container 1101 is configured to contain a biological sample therein. In some examples, container 1101 may be container 501 of the sample preservation and/or processing device of the present disclosure described with reference to FIG. 5 . That is, after the subject's biological sample is transferred to the container 501 of the sample preservation and/or processing device of the present disclosure, the processor cover 1102 is coupled to and seals the opening of the container 501 .

Processor lid 1102 may be configured to seal the opening of container 1101 . The processor cover may include at least one chamber containing at least one reagent necessary for processing the biological sample. In some examples, the processor cover may include two or more chambers, each chamber containing the same or different reagents therein. For example, the first chamber may be pre-filled with sample processing liquid. The sample processing solution and the biological sample are mixed in proportion to obtain the processed sample. The second chamber may be prefilled with reconstitution buffer. The third chamber may be pre-loaded with, for example, a PCR reaction solution, which contains components such as RT enzyme, taq enzyme, primers and probes required for the PCR reaction solution. In some cases, solid reagents, such as lyophilized powder or reagent pellets, may be pre-stored in the chamber. Reagents within the chamber of the processor lid can be released through the opening of the container into the opening.

In some examples, the chamber includes a pierceable element that interfaces with the opening of the container. The pierceable element is located at the lower end of the disposer cover (ie, the end that interfaces with the opening of the container) for sealing the at least one chamber of the disposer cover. In one example, the pierceable element may include a membrane. The at least one chamber of the processor cover may include a reagent release. When the reagent release member is activated, at least one reagent prestored in the corresponding chamber is released into the container. exist In one example, the agent release member includes a piercing member having a pointed end. The piercing member may be located within the at least one chamber. When the displacement of the piercing member in the direction toward the pierceable element exceeds a preset threshold (for example, exceeding the original distance from the tip of the piercing member to the pierceable element), the piercing member moves at least A portion of the chamber is pierced, allowing the reagents in the corresponding chamber to flow by gravity into the container below. In some examples, the agent release member may also include a drive member coupled to an end of the piercing member opposite the tip. The actuation component may be a button 1104. When force is applied to the key, the piercing member can be driven toward the pierceable element. The driving component may also include a spring for providing resistance to the driving component to prevent the driving component from being mistakenly pressed. For example, a predetermined biasing force provided by a spring needs to be overcome to drive the piercing member toward the pierceable element. In some cases, the keys are marked with symbols, such as 1, 2, 3..., to indicate to the user the order in which to operate the keys (i.e., the order in which the solution in the corresponding chamber is released into the container).

In some examples, the chamber includes a pressure-breakable element that interfaces with the opening of the container. The pressure breakable element is located at the lower end of the processor cover and is used to seal the at least one chamber of the processor cover. In one example, the pressure breakable element may include a membrane that breaks above a predetermined pressure. The at least one chamber of the processor cover may include a reagent release. When the reagent release member is activated, at least one reagent prestored in the corresponding chamber is released into the container. In one example, the reagent release member includes a pressure adjustment device, such as a piston rod and a piston coupled to one end of the piston rod. The piston is a tight fit against the inner wall of at least one chamber such that movement of the piston within the chamber causes a change in pressure within the chamber. When the pressure increase in the chamber exceeds the preset threshold, at least a portion of the pressure-breakable element is broken, so that the reagents in the corresponding chamber enter the container below under the action of gravity and pressure. In some examples, the reagent release may also include a drive component coupled to the piston rod. The actuation component may be a button. When force is applied to the key, the piston rod moves the piston toward the pressure-breaking element. The driving component may also include a spring for providing resistance to the driving component to prevent the driving component from being mistakenly pressed. For example, a predetermined biasing force provided by a spring needs to be overcome to drive the piston towards the pressure breakable element. In some examples, the pressure threshold that causes the pressure breakable element to break can be at least 1.1 times, 1.2 times, 1.4 times, 1.6 times, 1.8 times, 2.0 times, 2.2 times, 2.4 times, 2.6 times, 2.8 times, 3.0 times atmospheric pressure. times, 3.2 times, 3.4 times, 3.6 times, 3.8 times, 4.0 times, 4.2 times, 4.4 times, 4.6 times, 4.8 times, 5.0 times. The pressure threshold can be a value between any two of the above values.

The sample processing device of the present disclosure can be operated by an automated sample injector to realize automated processing of samples. The automated sampler may include a positioning component configured to accurately position the sample processing device of the present disclosure at a preset position of the operating platform of the sampler and prevent it from moving in the horizontal direction, and a pressing component, It is configured to compress the sample processing device of the present disclosure at a preset position of the operation platform of the sample injector to prevent it from moving in the vertical direction. The sample injector further includes a push rod mechanism, and the push rod mechanism may include one or more push rods for operating the reagent release member of the sample processing device. In one example, for each reagent release member of the sample processing device, a corresponding push rod can be provided, which contacts and pushes the corresponding reagent release member driven by the driving device, thereby sequentially releasing the reagents in the corresponding chamber. Release into the container of the sample processing device. In one example, only one push rod can be provided for multiple reagent release parts of the sample processing device, and the push rod sequentially contacts and pushes the corresponding reagent release parts driven by the driving device, thereby sequentially releasing the reagents in the corresponding chambers. into the container of the sample processing device.

Figure 13 is a flow chart depicting an exemplary method of processing a biological sample of the present disclosure. In process 1301, a sample processing device is provided that includes a container and a processor lid. In process 1302, a biological sample is placed within a container of a sample processing device. In process 1303, the disposer cover is coupled to the opening of the container. In process 1304, at least one reagent is released from the processor lid into the container.

In process 1301, the sample processing device provided may be the sample processing device of the present disclosure described with reference to FIGS. 11 and 12 . The container of the sample processing device has an opening at one end. The container is configured to hold a biological sample therein. The cover of the sample processing device is configured to seal the opening of the container and includes at least one chamber containing therein at least one reagent necessary for processing the biological sample. In some examples, the processor cover may include two or more chambers, each chamber containing the same or different reagents therein.

In process 1302, a biological sample is placed within the container through the opening of the container of the sample processing device. Subsequently, in process 1303, the closure is coupled to the opening of the container thereby sealing the opening.

In process 1304, at least one reagent is released into a container from at least one chamber of a processor cover of the sample processing device by activating the reagent release. In some examples, the chamber includes a pierceable element (eg, a pierceable membrane) that interfaces with the opening of the container, and the reagent release member includes a piercing member having a tip located within the chamber. When the displacement of the piercing member in the direction toward the pierceable element exceeds a preset threshold, the piercing member pierces at least a portion of the pierceable element, so that the reagent in the corresponding chamber flows into the lower chamber under the action of gravity. in the container. In other examples, the chamber includes a pressure breakable element (e.g., a membrane that breaks beyond a predetermined pressure) that interfaces with an opening of the container, and the reagent release member includes a pressure adjustment device (e.g., a sealant that is tightly fitted in at least one of the chambers). piston on the inner wall). When the piston is operated such that the pressure increase in the chamber exceeds the preset threshold, at least a portion of the pressure-breakable element is broken, thereby allowing the reagents in the chamber to enter the container below under the action of gravity and pressure.

The biological sample processed by the sample processing device of the present disclosure ultimately forms a reaction mixture. The reaction mixture may undergo further processing, such as a nucleic acid amplification reaction, within the container of the sample processing device of the present disclosure. The reaction mixture can be transferred to a reaction vessel and subjected to, for example, a nucleic acid amplification reaction in the reaction vessel. In one example, the container of the sample processing device of the present disclosure can be operated with a thermal cycler (eg, a PCR thermal cycler) to amplify the nucleic acid of interest in the sample and detect the amplification product. For example, the temperature of the container can be raised and lowered through a thermal cycler, thereby performing a polymerase chain reaction on the reaction mixture containing the biological sample in the container.

The target nucleic acid can be amplified in the container to generate an amplification product. The target nucleic acid can be target RNA or target DNA. In some embodiments, the target RNA is viral RNA. In some embodiments, the viral RNA may be pathogenic to the subject. Non-limiting examples of pathogenic viral RNA include human immunodeficiency virus I (HIV I), human immunodeficiency virus II (HIV II), orthomyxovirus, Ebola virus, dengue virus, influenza virus (e.g., H1N1, H3N2, H7N9, or H5N1), hepatitis virus, hepatitis A virus, hepatitis B virus, hepatitis C virus (e.g., RNA-HCV virus), hepatitis D virus, hepatitis E virus, hepatitis G virus, Epstein-Barr virus , mononucleosis virus, cytomegalovirus, SARS virus, West Nile virus, poliovirus, measles virus, or coronavirus (e.g., COVID-19). In the case where the target nucleic acid is target DNA, the target DNA may be any Types of DNA, including those described elsewhere herein. In some embodiments, the target DNA is viral DNA. In some embodiments, the viral DNA may be pathogenic to the subject. Non-limiting examples of DNA viruses include herpes simplex virus, variola virus, adenovirus (eg, adenovirus type 55, adenovirus type 7), or varicella virus (eg, fowlpox). In some cases, the target DNA can be bacterial DNA. The bacterial DNA may be from a bacterium that is pathogenic to the subject, for example, Mycobacterium tuberculosis, a bacterium known to cause tuberculosis. In some cases, the target DNA may be DNA from a pathogenic protozoan (eg, one or more Plasmodium-type protozoans that cause malaria).

Any type of nucleic acid amplification reaction known in the art can be used to amplify the target nucleic acid and generate an amplification product. Furthermore, amplification of nucleic acids can be linear, exponential, or a combination thereof. Amplification may be emulsion-based or may be non-emulsion-based. Non-limiting examples of nucleic acid amplification methods include reverse transcription, primer extension, polymerase chain reaction, ligase chain reaction, helicase-dependent amplification, asymmetric amplification, rolling circle amplification, and multiple displacement amplification (MDA ). In some embodiments, the amplification product can be DNA. Where target RNA is amplified, DNA can be obtained by reverse transcription of the RNA and subsequent DNA amplification can be used to generate an amplified DNA product. Amplified DNA products can indicate the presence of target RNA in the biological sample. Where DNA is amplified, any DNA amplification method known in the art may be used. Non-limiting examples of DNA amplification methods include polymerase chain reaction (PCR), variations of PCR (e.g., real-time PCR, allele-specific PCR, assembly PCR, asymmetric PCR, digital PCR, emulsion PCR, dial-out PCR (dial-out PCR), helicase-dependent PCR, nested PCR, hot-start PCR, inverse PCR, methylation-specific PCR, miniprimer PCR, multiplex PCR, nested PCR, overlap- Extension PCR, thermal asymmetric interlaced PCR, descending PCR) and ligase chain reaction (LCR). In some cases, DNA amplification is linear. In some cases, DNA amplification is exponential. In some cases, DNA amplification is accomplished using nested PCR, which improves the sensitivity of detecting the amplified DNA product.

In some cases, magnetic beads can be used to extract nucleic acids from biological samples in the container. Magnetic beads may be pre-stored in the nucleic acid extraction chamber. Magnetic beads adsorb nucleic acids in high-salt, low-PH solutions, and detach nucleic acids from the surface of magnetic beads in low-salt solutions, so magnetic beads can be used for biomass synthesis. Samples were extracted for nucleic acid extraction. The magnetic beads may be nanomagnetic beads, such as silicone hydroxyl magnetic beads or carboxyl magnetic beads. In one example, the magnetic beads are spherical particles composed of magnetic microspheres such as ferric oxide or ferric oxide and various materials such as silica containing active functional groups. The magnetic beads have instantaneous magnetic responsiveness, that is, superparamagnetism. Therefore, the magnetic beads can be positioned, guided and separated under the action of an external magnetic field.

The exemplary method of processing a biological sample of the present disclosure shown in Figure 13 can be performed by a computer system. The present disclosure also provides computer systems programmed to perform the methods of the present disclosure. Figure 14 illustrates a computer system 1401 programmed or otherwise configured to implement a sample collection method or sample processing method provided by the present disclosure. A computer system can regulate various aspects of the methods of the present disclosure. Computer system 1401 may be a user's electronic device or a computer system located remotely relative to the electronic device. The electronic device may be a mobile electronic device.

Computer system 1401 includes a central processing unit (CPU, also referred to herein as a "processor" and "computer processor") 1405, which may be a single-core or multi-core processor, or multiple processors for parallel processing. Computer system 1401 also includes a memory or memory location 1410 (e.g., random access memory, read only memory, flash memory), an electronic storage unit 1415 (e.g., a hard disk), a communication interface 1420 (e.g., a network) for communicating with one or more other systems. adapter) and peripheral devices 1425 such as cache, other memory, data storage, and/or electronic display adapters. The memory 1410, storage unit 1415, interface 1420 and peripheral devices 1425 communicate with the CPU 1405 through a communication bus (solid lines) such as the motherboard. The storage unit 1415 may be a data storage unit (or data storage library) for storing data. Computer system 1401 may be operatively coupled to a computer network ("network") 1430 via communication interface 1420. Network 1430 may be the Internet, the Internet and/or an extranet, or an intranet and/or extranet in communication with the Internet. In some cases, network 1430 is a telecommunications and/or data network. Network 1430 may include one or more computer servers that may enable distributed computing, such as cloud computing. In some cases, network 1430 may implement a peer-to-peer network with the help of computer system 1401 , which may enable devices coupled to computer system 1401 to act as clients or servers.

The CPU 1405 can execute a series of machine-readable instructions, which can be embodied in a program or software middle. Instructions may be stored in a memory location such as memory 1410. Instructions may be directed to CPU 1405, which may subsequently program or otherwise configure CPU 1405 to implement the methods of the present disclosure. Examples of operations performed by CPU 1405 may include fetch, decode, execute, and write back.

CPU 1405 may be part of a circuit, such as an integrated circuit. One or more other components of system 1401 may be included in the circuitry. In some cases, the circuit is an application specific integrated circuit (ASIC).

The storage unit 1415 may store files such as drivers, libraries, and saved programs. The storage unit 1415 may store user data such as user preferences and user programs. In some cases, computer system 1401 may include one or more additional data storage units located external to computer system 1401, such as on a remote server that communicates with computer system 1401 via an intranet or the Internet.

Computer system 1401 may communicate with one or more remote computer systems over network 1430. For example, computer system 1401 may communicate with a user's remote computer system. Examples of remote computer systems include personal computers (e.g. portable PCs), tablet PCs or tablet computers (e.g. Galaxy Tab), phone, smartphone (e.g. Support Android devices, ) or personal digital assistant. Users may access computer system 1401 through network 1430.

Methods as described herein may be implemented by machine (eg, computer processor) executable code stored on an electronic storage location (eg, memory 1410 or electronic storage unit 1415) of computer system 1401. Machine-executable or machine-readable code may be provided in the form of software. During use, the code may be executed by processor 1405. In some cases, code may be retrieved from storage unit 1415 and stored in memory 1410 for ready access by processor 1405. In some cases, electronic storage unit 1415 may be eliminated and machine-executable instructions stored in memory 1410.

The code may be precompiled and configured for use on a machine with a processor suitable for executing the code, or may be compiled during runtime. The code may be provided in a programming language that may be selected to enable execution of the code in a precompiled or as-compiled manner.

Aspects of the systems and methods provided herein (eg, computer system 1401) may be embodied in programming. Aspects of the technology may be considered a "product" or "artifact" in the form of machine (or processor) executable code and/or associated data, typically carried on or embodied in a type of machine-readable medium. The machine executable code may be stored on an electronic storage unit, such as a memory (eg, read-only memory, random access memory, flash memory) or a hard disk. "Storage" type media may include any or all of the tangible memory of a computer, processor, etc., or its related modules, such as various semiconductor memories, tape drives, disk drives, etc., which may provide non-transitory storage of software programming at any time. All or portions of the Software may sometimes be communicated over the Internet or various other telecommunications networks. For example, such communications may enable software to be loaded from one computer or processor to another, such as from a management server or host computer platform to an application server. Accordingly, another type of medium that may carry software components includes light waves, electrical waves, and electromagnetic waves, such as over physical interfaces between local devices over wired and optical landline networks and over various air-links. Light waves, radio waves and electromagnetic waves used. Physical components that carry such waves, such as wired or wireless links, optical links, etc., can also be considered media that carry software. As used herein, unless limited to non-transitory tangible "storage" media, terms such as computer or machine "readable medium" refer to any medium that participates in providing instructions to a processor for execution.

Accordingly, machine-readable media, such as computer executable code, can take many forms, including, but not limited to, tangible storage media, carrier wave media, or physical transmission media. Non-volatile storage media include, for example, optical disks or magnetic disks, such as any storage device in any computer, etc., which may be used to implement the databases shown in the drawings, etc. Volatile storage media includes dynamic memory, such as the main memory of such computer platforms. Tangible transmission media include coaxial cable; copper wire and fiber optics, including the wires that make up the buses within a computer system. Carrier transmission media may take the form of electrical or electromagnetic signals, or acoustic or light waves, such as those generated during radio frequency (RF) and infrared (IR) data communications. Thus, common forms of computer-readable media include, for example: floppy disks, floppy disks, hard drives, magnetic tape, any other magnetic media, CD-ROM, DVD or DVD-ROM, any other optical media, punched cardboard tapes, tapes with a hole pattern Any other physical storage media, RAM, ROM, PROM and EPROM, FLASH-EPROM, any other memory chip or cartridge, a carrier wave that transmits data or instructions, a cable or link that transmits such a carrier wave, or any other medium from which a computer can read programming code and/or data. Many of these forms of computer-readable media may be involved in carrying one or more sequences of one or more instructions to a processor for execution.

Computer system 1401 may include or be in communication with an electronic display 1435 that includes a user interface (UI) 1440 for providing, for example, assay conditions and protocols. Examples of UI include, but are not limited to, graphical user interfaces (GUIs) and web-based user interfaces.

While preferred embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Numerous variations, modifications and substitutions will now occur to those skilled in the art without departing from the present invention. It should be understood that various alternatives to the embodiments of the invention described herein may be used in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims (71)

A sample collection device including: a mouthpiece configured to engage at least a portion of the subject's mouth; a variable length member with a first end coupled to the mouthpiece member, the variable length member having a first state and a second state, in the first state, the length The variable length member has a first length along its longitudinal axis, and in said second state, said variable length member has a second length along said longitudinal axis, said second length being greater than said first length; as well as a sampling head coupled to a second end of the variable length member substantially opposite the first end along the longitudinal axis, the sampling head being configured to operate when the variable length member is at In the second state, the device is in contact with the collection site of the subject and collects a biological sample from the collection site. The sample collection device of claim 1, wherein no force is applied to the variable length member in the first state. The sample collection device of claim 1, wherein the variable length member is deformable when a force is applied to the variable length member. The sample collection device of claim 1, wherein the variable length member includes two or more deformable ribs. The sample collection device of claim 4, wherein the variable length member includes a rod extending between the two or more deformable ribs and the sampling head. The sample collection device of claim 5, wherein at least a portion of the rod is deformable when force is applied to the rod. The sample collection device of claim 1, wherein said sampling head is removable from said second end of said variable length member. The sample collection device of claim 1, wherein the sampling head is configured to retain the biological sample collected from the collection site of the subject. The sample collection device of claim 8, wherein the sampling head includes at least one brush. The sample collection device of claim 1, wherein the mouthpiece member is configured to be located outside the at least a portion of the mouth of the subject. The sample collection device of claim 10, wherein the at least a portion of the mouth includes the subject's lips. The sample collection device of claim 11 , wherein the mouthpiece member is sized to abut the lips when the variable length member is located within the mouth of the subject. . The sample collection device of claim 1, further comprising a handle coupled to the mouthpiece member on a side opposite the variable length member. The sample collection device of claim 1, wherein the collection site of the subject includes an oropharyngeal site. A method of collecting a biological sample from a subject, comprising: (a) providing a sample collection device comprising (i) a mouthpiece member, (ii) a variable length member, a first end of the variable length member being coupled to the mouthpiece member, wherein said variable length member is in a first state having a first length along its longitudinal axis, and (iii) a sampling head coupled to said variable length member along said longitudinal axis and said a second end substantially opposite the first end, wherein the mouthpiece member engages at least a portion of the subject's mouth; (b) Extending the variable length member with the mouthpiece member engaged with the at least a portion of the subject's mouth such that the variable length member changes from the first state to A second state, wherein in said second state said variable length member has a second length along said longitudinal axis, said second length being greater than said first length, wherein said variable length member has In the second state, the sampling head is in contact with the collection site of the subject to collect the biological sample of the subject from the collection site; and (c) Remove the mouthpiece member from the subject's mouth. 15. The method of claim 15, wherein extending the variable length member includes applying a force to the variable length member in a direction substantially perpendicular to the longitudinal axis. The method of claim 16, wherein the force is applied from at least the upper jaw, lower jaw, inner cheek, or a combination thereof of the subject. 15. The method of claim 15, wherein no force is applied to the variable length member in the first state. 15. The method of claim 15, wherein the variable length member is deformable when a force is applied to the variable length member. 15. The method of claim 15, wherein the variable length member includes two or more deformable ribs. 15. The method of claim 15, wherein the variable length member includes a rod extending between the two or more deformable ribs and the sampling head. 21. The method of claim 21, wherein at least a portion of the rod is deformable when force is applied to the rod. 15. The method of claim 15, wherein said sampling head is removable from said second end of said variable length member. The method of claim 15, wherein the sampling head is configured to retain the biological sample collected from the collection site of the subject. The method of claim 24, wherein said sampling head includes at least one brush. The method of claim 15, wherein the mouthpiece is configured to be located outside at least a portion of the subject's mouth. The method of claim 26, wherein the at least a portion of the mouth includes the subject's lips. 27. The method of claim 27, wherein the mouthpiece member is sized to abut the lips when the variable length member is located within the mouth of the subject. 15. The method of claim 15, wherein the sample collection device further includes a handle coupled to the mouthpiece member on a side opposite the variable length member. 16. The collection site of claim 15, wherein the collection site of the subject includes an oropharyngeal site. A test kit including: The sample collection device according to claim 1; and Optionally, instructions are provided to guide the user to use the sample collection device to collect a biological sample from the subject. A method of sample collection, including: (a) providing a sample collection device comprising (i) a mouthpiece member configured to engage at least a portion of a subject's mouth, (ii) a variable length member, A first end of the variable length member is coupled to the mouthpiece member, the variable length member has a first state and a second state, in the first state, the variable length member has an edge along the a first length of its longitudinal axis, the variable length member having a second length along the longitudinal axis in the second state, the second length being greater than the first length, and (iii) sampling a head coupled to a second end of the variable length member substantially opposite the first end along the longitudinal axis, the sampling head being configured to operate when the variable length member is in the The second state is in contact with the collection site of the subject and collects a biological sample from the collection site, wherein the sampling head includes the biological sample of the subject; and (b) Place the sampling head in the container. 32. The method of claim 32, wherein the container has an opening at one end, the container being configured to contain therein at least one reagent necessary for processing the biological sample. The method of claim 33, wherein the container includes a closure configured to seal the opening and includes a sample holder. 34. The method of claim 34, wherein the sample holder includes a material adapted to transfer the biological sample from the sampling head. The method of claim 34, wherein the sample holder is configured to hold at least a portion of the sampling head. A sample collection device including: a mouthpiece configured to engage at least a portion of the subject's mouth; A swab support coupled to the mouthpiece member and configured to support a swab, wherein the swab has a first position relative to the mouthpiece member and is different from the first A first position of the position, wherein the swab when in the second position is in contact with a collection site of the subject and collects a biological sample from the collection site. The sample collection device of claim 37, wherein at least a portion of the mouthpiece member is deformable. The sample collection device of claim 37, wherein the swab support is opposite The orientation of the mouthpiece member is changeable. 37. The sample collection device of claim 37, wherein the position of the swab relative to the swab support is variable. The sample collection device of claim 40, wherein the swab is coupled to the swab support through an elastic member. The sample collection device of claim 37, wherein the swab is coupled to the swab support by a buffer configured to regulate the collection of the subject by the swab. The force exerted by the part. 37. The sample collection device of claim 37, further comprising a handle coupled to the swab on a side opposite the collection site of the subject. The sample collection device of claim 37, wherein the mouthpiece member is configured to be located outside the at least a portion of the mouth of the subject. The sample collection device of claim 44, wherein the at least a portion of the mouth includes the subject's lips. The sample collection device of claim 37, further comprising a removable cover configured to cover at least the swab head of the swab. The sample collection device of claim 37, wherein the collection site of the subject includes an oropharyngeal site. A method of collecting a biological sample from a subject, comprising: (a) Provide a sample collection device, the sample collection device comprising (i) a mouthpiece member, (ii) a swab support member coupled to the mouthpiece member and configured to support the swab wherein the swab has a first position relative to the mouthpiece member, wherein the mouthpiece member engages at least a portion of the subject's mouth; (b) with the mouthpiece member engaged with the at least a portion of the subject's mouth, changing the swab from the first position to a third position relative to the mouthpiece member two positions, the second position being different from the first position, wherein the swab when in the second position is in contact with a collection site of the subject and collects a biological sample from the collection site; and (c) Remove the mouthpiece member from the subject's mouth. The method of claim 48, wherein said swab is moved from said first position to Changing to the second position includes changing the position of the swab relative to the swab support. 49. The method of claim 49, wherein changing the position of the swab relative to the swab support includes applying a force to the swab that is greater than a preset value. The method of claim 50, wherein the force greater than a preset value is applied by a handle coupled to the swab on a side opposite the collection site of the subject . 48. The method of claim 48, wherein changing the swab from the first position to the second position includes changing the orientation of the swab support relative to the mouthpiece member. The method of claim 52, wherein changing the orientation of the swab support relative to the mouthpiece member includes supporting the swab in a direction substantially perpendicular to a longitudinal axis of the swab support. The component exerts force. The method of claim 48, wherein the mouthpiece is configured to be located outside the at least a portion of the mouth of the subject. The method of claim 54, wherein the at least a portion of the mouth includes the subject's lips. A test kit including: The sample collection device of claim 37; and Optionally, instructions are provided to guide the user to use the sample collection device to collect biological samples. A sample processing device including: a container having an opening at one end, the container being configured to contain a biological sample therein; and A closure configured to seal the opening of the container and including at least one chamber containing at least one reagent necessary for processing the biological sample therein, the at least one chamber including a reagent release member , when the reagent release member is activated, the at least one reagent is released into the container. 57. The sample processing device of claim 57, wherein said chamber includes a pierceable element that interfaces with said opening of said container. The sample processing device of claim 58, wherein said pierceable element includes a membrane. The sample processing device of claim 57, wherein the reagent release member includes a driving member and a piercing member coupled to the driving member. The sample processing device of claim 60, wherein said piercing member is located within said chamber. The sample processing device of claim 60, wherein the driving member includes a button, and force is applied to the button to drive the piercing member toward the opening. 57. The sample processing device of claim 57, wherein said cover includes two or more said chambers, each of said chambers containing therein the same or different reagents from each other. A method of processing biological samples, comprising: (a) Provide a sample processing device, the sample processing device comprising (i) a container having an opening at one end, the container being configured to receive a biological sample therein, and (ii) a closure, the closure a lid configured to seal the opening of the container and include at least one chamber containing therein at least one reagent necessary for processing the biological sample; (b) placing the biological sample within the container through the opening; (c) coupling the closure to the opening of the container; and (d) activating a reagent release member to release said at least one reagent into said container. 64. The method of claim 64, wherein said chamber includes a pierceable element that interfaces with said opening of said container. The method of claim 65, wherein said pierceable element includes a membrane. 64. The method of claim 64, wherein the agent release member includes a drive member and a piercing member coupled to the drive member. 67. The method of claim 67, wherein said piercing member is located within said chamber. 67. The method of claim 67, wherein the driving member includes a button, and force is applied to the button to drive the piercing member toward the opening. 64. The method of claim 64, wherein said closure includes two or more said chambers, each of said chambers containing therein the same or different reagents from each other. A test kit including: The sample processing device of claim 57; and Optionally, instructions guide the user to process a biological sample using the sample processing device.