[go: up one dir, main page]
More Web Proxy on the site http://driver.im/

WO2024119245A1 - Méthodes destinées au diagnostic et au traitement d'une infection rhinosinusienne virale et bactérienne - Google Patents

Méthodes destinées au diagnostic et au traitement d'une infection rhinosinusienne virale et bactérienne Download PDF

Info

Publication number
WO2024119245A1
WO2024119245A1 PCT/AU2023/051279 AU2023051279W WO2024119245A1 WO 2024119245 A1 WO2024119245 A1 WO 2024119245A1 AU 2023051279 W AU2023051279 W AU 2023051279W WO 2024119245 A1 WO2024119245 A1 WO 2024119245A1
Authority
WO
WIPO (PCT)
Prior art keywords
biomarkers
sinonasal
protein
lnterleukin
necrosis factor
Prior art date
Application number
PCT/AU2023/051279
Other languages
English (en)
Inventor
El Din Rostom
Adam DAMRY
Jonathan Limpah
David Yen
Brian Wang
Tanya LUPANCU
Original Assignee
Diag-Nose Medical Pty Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Diag-Nose Medical Pty Ltd filed Critical Diag-Nose Medical Pty Ltd
Publication of WO2024119245A1 publication Critical patent/WO2024119245A1/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/112—Disease subtyping, staging or classification
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/14—Disorders of ear, nose or throat
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/26—Infectious diseases, e.g. generalised sepsis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/60—Complex ways of combining multiple protein biomarkers for diagnosis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/70—Mechanisms involved in disease identification
    • G01N2800/7095—Inflammation

Definitions

  • the present invention relates to methods and kits for the diagnosis of sinonasal inflammatory conditions caused by viral and bacterial infections. Furthermore, the present invention relates to methods for determining an appropriate treatment for a sinonasal inflammatory condition caused by a viral or bacterial infection and monitoring the response. The methods involve detecting biomarkers in sinonasal samples to assist in the differentiation between viral and bacterial infections and distinguish between disease states.
  • Sinonasal inflammatory conditions are characterised by the inflammation of mucous membranes that line the nasal cavity (rhinitis), the paranasal sinuses (sinusitis) or both (rhinosinusitis) and are classified according to the duration of symptoms including acute (less than 4 weeks), sub-acute (4-12 weeks) and chronic (over 12 weeks).
  • Inflammation of the mucous membranes may either lead to nasal drip or may prevent mucous drainage, thereby causing mucous obstruction.
  • Symptoms associated with sinonasal inflammatory conditions include swelling, pressure, and sinus pain.
  • Other symptoms commonly associated with sinonasal inflammatory conditions include sneezing, fever, headaches, loss of smell, sore throat, mucous discharge, and coughing.
  • Sinonasal inflammatory conditions are prevalent, whereby chronic rhinosinusitis affects around 10% of the population annually.
  • the most common causes of sinonasal inflammatory conditions are viral infections, bacterial infections, and allergic immune responses such as hay-fever. In some rare cases, particularly in people with weakened immune systems, a fungal infection can be the cause.
  • GP general practitioner
  • tests may be conducted to determine the cause. If an allergen is suspected to be the cause, an allergy test would be conducted. If viral or bacterial infections are suspected as being the cause, sampling techniques including nasopharyngeal swabs may be employed to collect samples from the nasal cavity for subsequent testing. However, typical sampling techniques are insufficient as the nasal cavity is colonised with an abundance of microbiota, therefore potentially resulting in falsely detecting commensal bacteria as pathogenic bacteria.
  • ENT Ear, Nose and Throat
  • ENT specialists perform nasal endoscopies to examine the sinonasal cavities using specialised tools including endoscopes and speculums.
  • Nasal endoscopy is performed to observe the presence of nasal polyps, swelling, and mucosal changes.
  • a CT scan may be performed in tandem to define the anatomy of the paranasal sinuses and observe any abnormalities.
  • ENT specialists may collect tissue culture samples from the middle meatus (MM), the cavity between the middle turbinate and lateral nasal wall for subsequent testing.
  • the microbiota of the MM has been found to be 80-90% representative of the microbiota in the sinuses and MM samples are less prone to being contaminated by commensal bacteria.
  • this modality requires endoscopic vision and is limited to ENT specialists or other trained personnel.
  • this modality is highly invasive, may cause significant discomfort, and may cause bleeding, leading to sample contamination.
  • Alternative diagnostic methods include collecting blood samples to measure the concentrations of one or more biomarkers associated with viral and bacterial infections.
  • Blood is collected via venepuncture, an invasive process which involves inserting a needle into a superficial vein to draw blood.
  • blood is an inadequate proxy for the nasal microenvironment as confounding factors due to comorbidities including cardiovascular disease, trauma, burns or surgical interventions may affect the accurate measurement of biomarkers, therefore affecting the diagnosis.
  • Blood biomarker levels may also be affected by drugs that act systemically. Additionally, the onset of fever can be required for blood biomarker tests which further limits the utility of blood tests.
  • the present invention relates to a method for diagnosing a sinonasal inflammatory condition in a patient caused by a viral or bacterial infection comprising measuring concentrations of a combination of biomarkers in a sinonasal sample collected from a patient and comparing the concentration of the biomarkers to a reference diagnostic value, wherein the biomarkers are selected from a panel of biomarkers including Human p-Defensin 2 (BD-2), C-reactive protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte- Colony Stimulating Factor (G-CSF), Granzyme B, lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-37 (IL- 37), Interferon gamma inducible protein-10 (IP-10), Matrix metalloproteinase-1 (MMP-1 ), Matrix
  • the biomarkers are RNA biomarkers. In other embodiments, the one or more biomarkers are protein biomarkers.
  • the biomarkers for detection of a viral cause of sinonasal inflammation include Human Myxovirus Resistance Protein 1 (MxA), Granzyme B, Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Serum Amyloid A (SAA), Tumor necrosis factor A (TNFa), Tumour Necrosis Factor- Related Apoptosis- Inducing Ligand (TRAIL), and Triggering receptor expressed on myeloid cells-2 (TREM- 2) and the biomarkers for detection of a bacterial cause of sinonasal inflammation include Human p-Defensin 2 (BD-2), C-Reactive Protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte colony stimulating factor (G-CSF), lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Myeloperoxida
  • MxA Human My
  • the present invention relates to a method for treating a sinonasal inflammatory condition in a patient comprising: determining whether the sinonasal inflammatory condition is caused by a viral infection or a bacterial infection based on measured concentrations of a combination of biomarkers; and administering a therapeutically effective amount of an antibiotic if the patient is identified as having a bacterial infection or administering an antiviral agent if the patient is identified as having a viral infection; wherein the biomarkers are selected from Human p-Defensin 2 (BD-2), C-reactive protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte- Colony Stimulating Factor (G-CSF), Granzyme B, lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-37 (IL-37), Interferon gamma inducible
  • BD-2 Human
  • the present invention relates to a method for monitoring a response of a patient diagnosed with a sinonasal inflammatory condition caused by a bacterial and/or viral infection to a treatment or therapy including: measuring concentrations of a combination of biomarkers in a sinonasal sample following the administration of the treatment or therapy; and comparing the measured concentrations of the biomarkers to reference diagnostic values; wherein the biomarkers are selected from Human p-Defensin 2 (BD-2), C-reactive protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte- Colony Stimulating Factor (G-CSF), Granzyme B, lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Matrix metalloproteinase-1
  • BD-2 Human p-
  • the present invention relates to a method for establishing reference diagnostic values to assist with bacterial and/or viral infection caused sinonasal inflammation profiling including: measuring concentrations of at least two biomarkers in sinonasal samples collected from a cohort of patients; wherein the biomarkers are selected from Human p-Defensin 2 (BD-2), C-reactive protein (CRP), Eotaxin-2, Eotaxin- 3, Granulocyte- Colony Stimulating Factor (G-CSF), Granzyme B, lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Matrix metalloproteinase-1 (MMP-1 ), Matrix metalloproteinase-8 (MMP-8), Myeloperoxidase (MPO), Human
  • BD-2 Human p
  • the cohort includes patients diagnosed with a sinonasal inflammatory condition caused by either a viral infection or a bacterial infection and not both.
  • the present invention relates to a kit for diagnosing sinonasal inflammatory conditions caused by viral or bacterial infections comprising reagents for measuring concentrations of a combination of biomarkers, wherein the biomarkers are selected from Human p-Defensin 2 (BD-2), C-reactive protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte- Colony Stimulating Factor (G-CSF), Granzyme B, lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Matrix metalloproteinase- 1 (MMP-1 ), Matrix metalloproteinase-8 (MMP-8), Myeloperoxidase (MPO), Human Myxovirus Resistance protein 1 (MxA), Neu
  • the reagents are antibodies. In other embodiments, the reagents are oligonucleotide probes.
  • the biomarkers for detection of a viral cause of sinonasal inflammation include Human Myxovirus Resistance Protein 1 (MxA), Granzyme B, Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Serum Amyloid A (SAA), Tumor necrosis factor A (TNFa), Tumour Necrosis Factor- Related Apoptosis- Inducing Ligand (TRAIL), and Triggering receptor expressed on myeloid cells-2 (TREM- 2) and the biomarkers for detection of a bacterial cause of sinonasal inflammation include Human p-Defensin 2 (BD-2), C-Reactive Protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte colony stimulating factor (G-CSF), lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), Interleukin-8 (IL-8), Interleukin-10 (IL-10), Myeloperoxidase (MxA), Granzyme
  • the combination of biomarkers includes any one or more of lnterleukin-3 (IL-3), lnterleukin-5 (IL-5) Matrix metalloproteinase-8 (MMP-8) and Triggering receptor expressed on myeloid cells-1 (TREM-1 ) and any one or more of Triggering receptor expressed on myeloid cells 2 (TREM-2), Serum amyloid A (SAA), Granzyme B, Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Tumor necrosis factor A (TNFa), and Tumour Necrosis Factor- Related Apoptosis- Inducing Ligand (TRAIL).
  • IL-3 lnterleukin-3
  • IL-5 IL-5
  • MMP-8 Matrix metalloproteinase-8
  • TRM-1 Triggering receptor expressed on myeloid cells-1
  • TAM-2 Triggering receptor expressed on myeloid cells 2
  • SAA Serum amyloid A
  • the combination of biomarkers includes any one or more of Granulocyte colony stimulating factor (G-CSF) and Myeloperoxidase (MPO), and any one or more of Serum amyloid A (SAA), Granzyme B, Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Tumor necrosis factor A (TNFa), Tumour Necrosis Factor- Related Apoptosis-Inducing Ligand (TRAIL), and Human Myxovirus Resistance Protein 1 (MxA).
  • G-CSF Granulocyte colony stimulating factor
  • MPO Myeloperoxidase
  • SAA Serum amyloid A
  • Granzyme B Granzyme B
  • IL-37 Interleukin-37
  • IP-10 Interferon gamma inducible protein-10
  • TNFa Tumor necrosis factor A
  • TRAIL Tumour Necrosis Factor- Related Apoptosis-Inducing Ligand
  • the combination of biomarkers includes any one or more of Eotaxin-3 and Matrix metalloproteinase- 1 (MMP-1 ) and any one or more of Serum amyloid A (SAA), Granzyme B, Interferon gamma inducible protein-10 (IP-10), and T umor necrosis factor A (TNFa).
  • MMP-1 Matrix metalloproteinase- 1
  • SAA Serum amyloid A
  • IP-10 Interferon gamma inducible protein-10
  • TNFa T umor necrosis factor A
  • the combination of biomarkers includes any one or more of Human p-Defensin 2 (BD-2) and Neutrophil Elastase (NE) and any one or more of Human Myxovirus Resistance Protein 1 (MxA), Tumour Necrosis Factor- Related Apoptosis- Inducing Ligand (TRAIL), and Interferon gamma inducible protein-10 (IP-10).
  • BD-2 Human p-Defensin 2
  • NE Neutrophil Elastase
  • MxA Human Myxovirus Resistance Protein 1
  • TRAIL Tumour Necrosis Factor- Related Apoptosis- Inducing Ligand
  • IP-10 Interferon gamma inducible protein-10
  • the combination of biomarkers includes Eotaxin-2 and any one or more of Serum amyloid A (SAA), Granzyme B, Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Tumor necrosis factor A (TNFa), and Tumour Necrosis Factor- Related Apoptosis-Inducing Ligand (TRAIL).
  • SAA Serum amyloid A
  • IL-37 Interleukin-37
  • IP-10 Interferon gamma inducible protein-10
  • TNFa Tumor necrosis factor A
  • TRAIL Tumour Necrosis Factor- Related Apoptosis-Inducing Ligand
  • the combination of biomarkers includes Human Myxovirus Resistance Protein 1 (MxA) and any one or more of Triggering receptor expressed on myeloid cells 2 (TREM-2) and Serum amyloid A (SAA).
  • MxA Human Myxovirus Resistance Protein 1
  • TREM-2 Triggering receptor expressed on myeloid cells 2
  • SAA Serum amyloid A
  • the combination of biomarkers includes lnterleukin-8 (IL-8) and any one or more of Triggering receptor expressed on myeloid cells 2 (TREM-2), Serum amyloid A (SAA), Granzyme B, Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Tumor necrosis factor A (TNFa), Tumour Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL), and Human Myxovirus Resistance Protein 1 (MxA).
  • TNFa Tumor necrosis factor A
  • TRAIL Tumour Necrosis Factor-Related Apoptosis-Inducing Ligand
  • MxA Human Myxovirus Resistance Protein 1
  • the combination of biomarkers includes C-reactive protein (CRP) and any one or more of Interferon gamma inducible protein-10 (IP-10) and Human Myxovirus Resistance Protein 1 (MxA).
  • CRP C-reactive protein
  • IP-10 Interferon gamma inducible protein-10
  • MxA Human Myxovirus Resistance Protein 1
  • the combination of biomarkers is Interleukin-10 and Tumour Necrosis Factor- Related Apoptosis-Inducing Ligand (TRAIL).
  • TRAIL Tumour Necrosis Factor- Related Apoptosis-Inducing Ligand
  • the sinonasal sample is sinonasal secretions, sinonasal aspirates or tissue cultures.
  • the one or more biomarkers are RNA biomarkers.
  • the one or more biomarkers are protein biomarkers.
  • kits for diagnosing sinonasal inflammatory conditions caused by viral or bacterial infections adapted for use with any one or more of multiplexing instruments, antibody/aptamer binding immunoassays, point- of-care microfluidic devices and lateral flow assays.
  • Figure 1 illustrates the distribution of BD-2 protein concentrations measured across patient groups in Study 1 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 2 illustrates an ROC curve for BD-2 derived from the data of Figure 1 .
  • Figure 3 illustrates the distribution of CRP protein concentrations measured across patient groups in Study 1 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 4 illustrates an ROC curve for CRP derived from the data of Figure 3.
  • Figure 5 illustrates the distribution of G-CSF protein concentrations measured across patient groups in Study 1 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 6 illustrates an ROC curve for G-CSF derived from the data of Figure 5.
  • Figure 7 illustrates the distribution of IL-8 protein concentrations measured across patient groups in Study 1 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 8 illustrates an ROC curve for IL-8 derived from the data of Figure 7.
  • Figure 9 illustrates the distribution of MPO protein concentrations measured across patient groups in Study 1 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 10 illustrates an ROC curve for MPO derived from the data of Figure 9.
  • Figure 11 illustrates the distribution of NE protein concentrations measured across patient groups in Study 1 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 12 illustrates an ROC curve for NE derived from the data of Figure 11 .
  • Figure 13 illustrates the distribution of SAA protein concentrations measured across patient groups in Study 1 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 14 illustrates an ROC curve for SAA derived from the data of Figure 13.
  • Figure 15 illustrates the distribution of TREM-2 protein concentrations measured across patient groups in Study 1 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 16 illustrates an ROC curve for TREM-2 derived from the data of Figure 15.
  • Figure 17 illustrates the distribution of IP-10 protein concentrations measured across patient groups in Study 1 according to the experimental method described herein below. Positive corresponds to viral patient group, Challenge corresponds to bacterial patient group, and Negative corresponds to Healthy patient group.
  • Figure 18 illustrates an ROC curve for IP-10 derived from the data of Figure 17.
  • Figure 19 illustrates the distribution of MxA protein concentrations measured across patient groups in Study 1 according to the experimental method described herein below. Positive corresponds to viral patient group, Challenge corresponds to bacterial patient group, and Negative corresponds to Healthy patient group.
  • Figure 20 illustrates an ROC curve for MxA obtained from the data of Figure 19.
  • Figure 21 illustrates the distribution of TRAIL protein concentrations measured across patient groups in Study 1 according to the experimental method described herein below. Positive corresponds to viral patient group, Challenge corresponds to bacterial patient group, and Negative corresponds to Healthy patient group.
  • Figure 22 illustrates an ROC curve for TRAIL obtained from the data of Figure 21.
  • Figure 23 comprises a table, identified as Table 1 , which includes cut off values and associated sensitivity and specificity values for diagnosing a bacterial infection or viral infection in Study 1 using the bacterial biomarkers Human p-Defensin 2 (BD-2), C- Reactive Protein (CRP), Granulocyte colony stimulating factor (G-CSF), lnterleukin-8 (IL- 8), Myeloperoxidase (MPO), Neutrophil Elastase (NE), Serum amyloid A (SAA), and Triggering receptor expressed on myeloid cells 2 (TREM-2), and viral biomarkers Interferon gamma inducible protein-10 (IP-10), Human Myxovirus Resistance Protein 1 (MxA), and Tumour Necrosis Factor- Related Apoptosis-Inducing Ligand (TRAIL).
  • BD-2 Human p-Defensin 2
  • CRP C- Reactive Protein
  • G-CSF Granulocyte colony stimulating factor
  • IL- 8 lnterleukin
  • Figure 24 comprises a table, identified as Table 2, which includes cut off values and associated sensitivity and specificity data for diagnosing a bacterial infection or viral infection in Study 1 using combinations of biomarkers selected from Human Myxovirus Resistance Protein 1 (MxA), Neutrophil Elastase (NE), Interferon gamma inducible protein-10 (IP-10), Human p-Defensin 2 (BD-2), lnterleukin-8 (IL-8), Myeloperoxidase (MPO), C-Reactive Protein (CRP), Tumour Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL), Triggering receptor expressed on myeloid cells 2 (TREM-2); Granulocyte colony stimulating factor (G-CSF) and Serum amyloid A (SAA).
  • MxA Human Myxovirus Resistance Protein 1
  • NE Neutrophil Elastase
  • IP-10 Interferon gamma inducible protein-10
  • IP-10 Human p-Defensin 2
  • Figure 25 illustrates the distribution of CRP protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 26 illustrates an ROC curve for CRP derived from the data of Figure 25.
  • Figure 27 illustrates the distribution of Eotaxin-2 protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 28 illustrates an ROC curve for Eotaxin-2 derived from the data of Figure 27.
  • Figure 29 illustrates the distribution of Eotaxin-3 protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 30 illustrates an ROC curve for Eotaxin-3 derived from the data of Figure 29.
  • Figure 31 illustrates the distribution of G-CSF protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 32 illustrates an ROC curve for G-CSF derived from the data of Figure 31.
  • Figure 33 illustrates the distribution of IL-3 protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 34 illustrates an ROC curve for IL-3 derived from the data of Figure 33.
  • Figure 35 illustrates the distribution of IL-5 protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 36 illustrates an ROC curve for IL-5 derived from the data of Figure 35.
  • Figure 37 illustrates the distribution of IL-8 protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 38 illustrates an ROC curve for IL-8 derived from the data of Figure 37.
  • Figure 39 illustrates the distribution of IL-10 protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 40 illustrates an ROC curve for IL-10 derived from the data of Figure 39.
  • Figure 41 illustrates the distribution of MMP-1 protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 42 illustrates an ROC curve for MMP-1 derived from the data of Figure 41.
  • Figure 43 illustrates the distribution of MMP-8 protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 44 illustrates an ROC curve for MMP-8 derived from the data of Figure 43.
  • Figure 45 illustrates the distribution of MPO protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 46 illustrates an ROC curve for MPO derived from the data of Figure 45.
  • Figure 47 illustrates the distribution of TREM-1 protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 48 illustrates an ROC curve for TREM-1 derived from the data of Figure 47.
  • Figure 49 illustrates the distribution of Granzyme B protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to viral patient group, Challenge corresponds to bacterial patient group, and Negative corresponds to Healthy patient group.
  • Figure 50 illustrates an ROC curve for Granzyme B derived from the data of Figure 49.
  • Figure 51 illustrates the distribution of IL-37 protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to viral patient group, Challenge corresponds to bacterial patient group, and Negative corresponds to Healthy patient group.
  • Figure 52 illustrates an ROC curve for IL-37 derived from the data of Figure 50.
  • Figure 53 illustrates the distribution of IP-10 protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to viral patient group, Challenge corresponds to bacterial patient group, and Negative corresponds to Healthy patient group.
  • Figure 54 illustrates an ROC curve for IP-10 derived from the data of Figure 53.
  • Figure 55 illustrates the distribution of SAA protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to viral patient group, Challenge corresponds to bacterial patient group, and Negative corresponds to Healthy patient group.
  • Figure 56 illustrates an ROC curve for SAA derived from the data of Figure 55.
  • Figure 57 illustrates the distribution of TNFa protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to viral patient group, Challenge corresponds to bacterial patient group, and Negative corresponds to Healthy patient group. [0095]
  • Figure 58 illustrates an ROC curve for TNFa derived from the data of Figure 57.
  • Figure 59 illustrates the distribution of TRAIL protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to viral patient group, Challenge corresponds to bacterial patient group, and Negative corresponds to Healthy patient group.
  • Figure 60 illustrates an ROC curve for TRAIL derived from the data of Figure 59.
  • Figure 61 illustrates the distribution of Granzyme B protein concentrations measured across patient groups in Study 2 according to the experimental method described herein below. Positive corresponds to viral patient group, Challenge corresponds to bacterial patient group, and Negative corresponds to Healthy patient group.
  • Figure 62 illustrates an ROC curve for Granzyme B derived from the data of Figure 61.
  • Figure 63 comprises a table, identified as Table 3, which includes cut off values and associated sensitivity and specificity values for diagnosing a bacterial infection or viral infection in Study 2 using the bacterial biomarkers Human ⁇ - C-Reactive Protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte colony stimulating factor (G-CSF), Interleukin- 3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Matrix metalloproteinase- 1 (MMP-1 ), Matrix metalloproteinase-8 (MMP-8), Myeloperoxidase (MPO), and Triggering receptor expressed on myeloid cells 1 (TREM-1 ), and viral biomarkers Granzyme B, Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Serum Amyloid A (SAA), Tumor necrosis factor A (TNFa), Tumor
  • Figure 64 comprises a table, identified as Table 4, which includes cut off values and associated sensitivity and specificity data for diagnosing a bacterial infection or viral infection in Study 2 using combinations of biomarkers selected from C-reactive protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte- Colony Stimulating Factor (G-CSF), Granzyme B, lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Matrix metalloproteinase- 1 (MMP-1 ), Matrix metalloproteinase-8 (MMP-8), Myeloperoxidase (MPO), Serum Amyloid A (SAA), Tumor necrosis factor A (TNFa), Tumor necrosis factor- related apoptosis-inducing ligand
  • Figure 65 illustrates the distribution of BD-2 protein concentrations measured across patient groups in Study 3 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 66 illustrates an ROC curve for BD-2 derived from the data of Figure 65.
  • Figure 67 illustrates the distribution of CRP protein concentrations measured across patient groups in Study 3 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 68 illustrates an ROC curve for CRP derived from the data of Figure 67.
  • Figure 69 illustrates the distribution of G-CSF protein concentrations measured across patient groups in Study 3 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 70 illustrates an ROC curve for G-CSF derived from the data of Figure 69.
  • Figure 71 illustrates the distribution of IL-8 protein concentrations measured across patient groups in Study 3 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 72 illustrates an ROC curve for IL-8 derived from the data of Figure 71 .
  • Figure 73 illustrates the distribution of MPO protein concentrations measured across patient groups in Study 3 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 74 illustrates an ROC curve for MPO derived from the data of Figure 73.
  • Figure 75 illustrates the distribution of NE protein concentrations measured across patient groups in Study 3 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 76 illustrates an ROC curve for NE derived from the data of Figure 75.
  • Figure 77 illustrates the distribution of SAA protein concentrations measured across patient groups in Study 3 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group.
  • Figure 78 illustrates an ROC curve for SAA derived from the data of Figure 77.
  • Figure 79 illustrates the distribution of TREM-2 protein concentrations measured across patient groups in Study 3 according to the experimental method described herein below. Positive corresponds to bacterial patient group, Challenge corresponds to viral patient group, and Negative corresponds to Healthy patient group. [00118]
  • Figure 80 illustrates an ROC curve for TREM-2 derived from the data of Figure 79.
  • Figure 81 illustrates the distribution of IP-10 protein concentrations measured across patient groups in Study 3 according to the experimental method described herein below. Positive corresponds to viral patient group, Challenge corresponds to bacterial patient group, and Negative corresponds to Healthy patient group.
  • Figure 82 illustrates an ROC curve for IP-10 derived from the data of Figure 81 .
  • Figure 83 illustrates the distribution of MxA protein concentrations measured across patient groups in Study 3 according to the experimental method described herein below. Positive corresponds to viral patient group, Challenge corresponds to bacterial patient group, and Negative corresponds to Healthy patient group.
  • Figure 84 illustrates an ROC curve for MxA obtained from the data of Figure 83.
  • Figure 85 illustrates the distribution of TRAIL protein concentrations measured across patient groups in Study 3 according to the experimental method described herein below. Positive corresponds to viral patient group, Challenge corresponds to bacterial patient group, and Negative corresponds to Healthy patient group.
  • Figure 86 illustrates an ROC curve for TRAIL obtained from the data of Figure 85.
  • Figure 87 comprises a table, identified as Table 5, which includes cut off values and associated sensitivity and specificity values for diagnosing a bacterial infection or viral infection in Study 3 using the bacterial biomarkers Human p-Defensin 2 (BD-2), C- Reactive Protein (CRP), Granulocyte colony stimulating factor (G-CSF), lnterleukin-8 (IL- 8), Myeloperoxidase (MPO), Neutrophil Elastase (NE), Serum amyloid A (SAA), and Triggering receptor expressed on myeloid cells 2 (TREM-2), and viral biomarkers Interferon gamma inducible protein-10 (IP-10), Human Myxovirus Resistance Protein 1 (MxA), and Tumour Necrosis Factor- Related Apoptosis-Inducing Ligand (TRAIL).
  • BD-2 Human p-Defensin 2
  • CRP C- Reactive Protein
  • G-CSF Granulocyte colony stimulating factor
  • IL- 8 lnterleukin-8
  • Figure 88 comprises a table, identified as Table 6, which includes cut off values and associated sensitivity and specificity data for diagnosing a bacterial infection or viral infection in Study 3 using combinations of biomarkers selected from Human Myxovirus Resistance Protein 1 (MxA), Neutrophil Elastase (NE), Interferon gamma inducible protein-10 (IP-10), Human p-Defensin 2 (BD-2), lnterleukin-8 (IL-8), Myeloperoxidase (MPO), C-Reactive Protein (CRP), Tumour Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL), Triggering receptor expressed on myeloid cells 2 (TREM-2); Granulocyte colony stimulating factor (G-CSF) and Serum amyloid A (SAA).
  • MxA Human Myxovirus Resistance Protein 1
  • NE Neutrophil Elastase
  • IP-10 Interferon gamma inducible protein-10
  • IP-10 Human p-Defensin 2
  • biomarker refers to a biological characteristic that is a quantitative indicator of biological processes, pathological processes, and pharmacological responses. Biomarkers are measured in biological samples such as bodily fluids and encompass genes, proteins, nucleic acids, fragments of nucleic acids, lipids, metabolites, carbohydrates, and hormones.
  • Biomarkers referred to herein include Human p-Defensin 2 (BD-2), C-reactive protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte- Colony Stimulating Factor (G-CSF), Granzyme B, lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Matrix metalloproteinase- 1 (MMP-1 ), Matrix metalloproteinase-8 (MMP-8), Myeloperoxidase (MPO), Human Myxovirus Resistance protein 1 (MxA), Neutrophil Elastase (NE), Serum Amyloid A (SAA), Tumor necrosis factor A (TNFa), Tumor necrosis factor-related apoptosis-inducing ligand (
  • the present invention relates to methods and kits for the diagnosis and treatment of sinonasal inflammatory conditions caused by viral and bacterial infections.
  • the methods include diagnosing sinonasal inflammatory conditions caused by viral and bacterial infections; determining an appropriate treatment for a sinonasal inflammatory condition caused by a viral or bacterial infection; and monitoring a patient’s response to treatment.
  • Each of the methods involve a) collecting sinonasal samples from a patient; b) detecting and measuring the concentration of biomarkers in the sinonasal sample associated with viral infections and bacterial infections; and c) comparing the measured concentrations to reference diagnostic values.
  • the present invention further relates to a method of establishing reference diagnostic values to assist with sinonasal inflammatory profiling.
  • the methods disclosed herein describe combinations of biomarkers detected in sinonasal samples for the accurate diagnosis of sinonasal inflammatory conditions that do not require invasive or site-specific sampling.
  • the present invention relates to a method for the accurate diagnosis of sinonasal inflammatory conditions caused by viral and bacterial infections.
  • the method comprises a) collecting sinonasal samples from a patient; b) detecting and measuring the concentration of biomarkers in the biological sample that are associated with viral infections and bacterial infections; and c) analysing and comparing the measured concentrations of the biomarkers to reference diagnostic values.
  • Sinonasal samples include sinonasal secretions, sinonasal aspirates and sinonasal tissue cultures.
  • the sinonasal sample is collected by forced exhalation or absorptive strips.
  • the dilution ratio of the sinonasal sample is a standardised dilution ratio for accurate testing.
  • the invention disclosed herein includes application of a sinonasal sample for the detection of one or more biomarkers that are upregulated in a patient exhibiting sinonasal inflammation caused by a viral or a bacterial infection.
  • the biomarkers detected and measured are selected from a protein biomarker panel comprising Human p-Defensin 2 (BD-2), C-reactive protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte- Colony Stimulating Factor (G-CSF), Granzyme B, lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Matrix metalloproteinase- 1 (MMP-1 ), Matrix metalloproteinase-8 (MMP-8), Myeloperoxidase (MPO), Human Myxovirus Resistance protein 1 (MxA), Neutrophil Elastase (NE), Serum Amyloid A (SAA), Tumor necrosis factor A (TNFa), Tumor necrosis factor-
  • BD-2 Human p
  • Biomarkers for detection of a viral cause of sinonasal inflammation include Human Myxovirus Resistance Protein 1 (MxA), Granzyme B, Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Serum Amyloid A (SAA), Tumor necrosis factor A (TNFa), Tumour Necrosis Factor- Related Apoptosis-Inducing Ligand (TRAIL), and Triggering receptor expressed on myeloid cells-2 (TREM-2).
  • MxA Human Myxovirus Resistance Protein 1
  • IL-37 Interleukin-37
  • IP-10 Interferon gamma inducible protein-10
  • SAA Serum Amyloid A
  • TNFa Tumor necrosis factor A
  • TRAIL Tumour Necrosis Factor- Related Apoptosis-Inducing Ligand
  • TAM-2 Triggering receptor expressed on myeloid cells-2
  • Biomarkers for detection of a bacterial cause of sinonasal inflammation include Human p-Defensin 2 (BD-2), C-Reactive Protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte colony stimulating factor (G-CSF), lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Myeloperoxidase (MPO), Matrix metalloproteinase- 1 (MMP-1 ), Matrix metalloproteinase-8 (MMP-8), Neutrophil Elastase (NE), and Serum amyloid A (SAA), Triggering receptor expressed on myeloid cells-1 (TREM-1 ), and Triggering receptor expressed on myeloid cells 2 (TREM-2).
  • BD-2 Human p-Defensin 2
  • CRP C-Reactive Protein
  • Eotaxin-2 Eotaxin-3
  • G-CSF Gran
  • the biomarkers are detected and measured using multiplexing kits such as LUMINEX multiplex, Meso Scale Discovery (MSD), Quanterix and Olink.
  • the biomarkers are detected and measured using ELISA, Western Blot, or flow cytometry.
  • point-of-care systems such as microfluidic chips or multiplexing lateral flow test strips are used to the detect and measure the biomarkers in vitro.
  • other techniques can be used to detect and measure biomarker concentrations.
  • Figure 23, Figure 63, and Figure 87 contains Table 1 , Table 3, and Table 5, respectively. These tables include cut off values and associated sensitivity and specificity values of biomarkers for bacterial infection or viral infection obtained from data derived from samples obtained from different studies according to the experimental method described herein below ( Figure 23 from Study 1 , Figure 62 from Study 2, and Figure 87 from Study 3).
  • the tables illustrate the cut-off concentrations of biomarkers in samples diluted at a specified dilution ratio or undiluted samples, wherein at Min, Peak, and Max cutoff concentrations, the Min Peak, and Max specificity and sensitivity are specified.
  • the tables also set out additional data for each of the biomarkers, including accuracy, area under the curve (AUC), 95% confidence interval, and P-value.
  • the cut-off concentrations of CRP in an undiluted sample wherein at a minimum cut-off concentration of 160,000, specificity is 76% and sensitivity is 100% for bacterial infection, and wherein at a maximum cut-off concentration of 8,000,000 ng/ml, specificity is 100% and sensitivity is 33% for bacterial infection, and wherein at a peak cut-off concentration of 600,000 ng/ml, specificity is 97% and sensitivity is 100% for bacterial infection. That is, 1 ,600 to 80,000 ng/ml with a peak of 6,000 ng/ml in a sample with a dilution ratio of 1 : 100.
  • the cutoff concentrations in Figures 23, 63, and 87 also represent the reference diagnostic range thresholds identified in the different studies.
  • a concentration of p- Defensin 2 within a range of 7,600 to 30,000 ng/ml is indicative of bacterial infection.
  • a concentration of CRP within a range of 160,000 ng/ml to 8,000,000 ng/ml is indicative of bacterial infection.
  • a concentration of G-CSF within a range of 4,300 to 50,000 ng/ml is indicative of bacterial infection.
  • a concentration of IL-8 within a range of 35,000 to 800,000 ng/ml is indicative of bacterial infection.
  • a concentration of MPO within a range of 20,000,000 to 200,000,000 ng/ml is indicative of bacterial infection.
  • a concentration of neutrophil elastase within a range of 23,000 to 720,000 ng/ml is indicative of bacterial infection.
  • a concentration of SAA within a range of 45,000 to 300,000 ng/ml is indicative of bacterial infection.
  • a concentration of TREM-2 within a range of 3,000 to 360,000 ng/ml is indicative of bacterial infection.
  • a concentration of IP-10 within a range of 50 to 800 ng/ml is indicative of viral infection.
  • a concentration of MxA within a range of 0.6 to 320 ng/ml is indicative of viral infection.
  • a concentration of TRAIL within a range of 3,000 to 22,000 ng/ml is indicative of viral infection.
  • the present invention relates to determining an appropriate treatment for a patient diagnosed with a sinonasal inflammatory condition caused by a viral or bacterial infection.
  • the method comprises: a) collecting a biological sample from a patient; b) detecting and measuring the concentration of biomarkers in the biological sample that are associated with viral infections and bacterial infections; c) analysing and comparing the measured concentrations of the biomarkers to reference diagnostic values; and d) administering a therapeutically effective amount of an antibiotic if the patient is diagnosed with a sinonasal inflammatory condition caused by a bacterial infection or administering a therapeutically effective amount of an antiviral if the patient is diagnosed with a sinonasal inflammatory condition caused by a virus.
  • Sinonasal samples include sinonasal secretions, sinonasal aspirates and sinonasal tissue cultures.
  • the sinonasal sample is collected by forced exhalation or absorptive strips.
  • the volume of the sinonasal sample is a standardised sample volume consistent with volumes required for accurate testing.
  • the biomarkers detected and measured are selected from a protein biomarker panel comprising Human p-Defensin 2 (BD-2), C-reactive protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte- Colony Stimulating Factor (G-CSF), Granzyme B, lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Matrix metalloproteinase- 1 (MMP-1), Matrix metalloproteinase-8 (MMP-8), Myeloperoxidase (MPO), Human Myxovirus Resistance protein 1 (MxA), Neutrophil Elastase (NE), Serum Amyloid A (SAA), Tumor necrosis factor A (TNFa), Tumor necrosis factor-related
  • BD-2 Human p
  • the biomarkers are detected and measured using multiplexing kits such as LUMINEX multiplex, Meso Scale Discovery (MSD), Quanterix and Olink.
  • the biomarkers are detected and measured using ELISA, Western Blot, or flow cytometry.
  • point-of-care systems such as microfluidic chips or multiplexing lateral flow test strips are used to detect and measure the biomarkers.
  • other techniques can be used to detect and measure biomarker concentrations.
  • the invention relates to a method for monitoring the response of a patient diagnosed with a sinonasal inflammatory condition caused by a viral or bacterial infection and administered a therapy or treatment.
  • the method comprises: a) collecting a biological sample from a patient; b) measuring the concentration of biomarkers in the biological sample that are associated with viral infections and/or bacterial infections; and c) analysing and comparing the measured concentrations of the biomarkers to reference diagnostic values.
  • Sinonasal samples include sinonasal secretions, sinonasal aspirates and sinonasal tissue cultures.
  • the sinonasal sample is collected by forced exhalation or absorptive strips.
  • the volume of the sinonasal sample is a standardised sample volume consistent with volumes required for accurate testing.
  • the biomarkers detected and measured are selected from a protein biomarker panel comprising Human p-Defensin 2 (BD-2), C-reactive protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte- Colony Stimulating Factor (G-CSF), Granzyme B, lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Matrix metalloproteinase- 1 (MMP-1 ), Matrix metalloproteinase-8 (MMP-8), Myeloperoxidase (MPO), Human Myxovirus Resistance protein 1 (MxA), Neutrophil Elastase (NE), Serum Amyloid A (SAA), Tumor necrosis factor A (TNFa), Tumor necrosis factor-
  • BD-2 Human p
  • the biomarkers are detected and measured using multiplexing instruments comprising bead or planar arrays such as LUMINEX and Quanterix instruments, electrochemiluminescence such as Meso Scale Discovery (MSD) arrays, proximity extension assays such as Olink, microfluidic arrays, homogenous or heterogenous immunoassay analysers such as Olympus AU400 and liquid chromatography-mass spectrometry (LC-MS).
  • multiplexing instruments comprising bead or planar arrays such as LUMINEX and Quanterix instruments, electrochemiluminescence such as Meso Scale Discovery (MSD) arrays, proximity extension assays such as Olink, microfluidic arrays, homogenous or heterogenous immunoassay analysers such as Olympus AU400 and liquid chromatography-mass spectrometry (LC-MS).
  • multiplexing instruments comprising bead or planar arrays such as LUMINEX and Quanterix instruments, electrochemiluminescence such as Meso Scal
  • the biomarkers are detected and measured using ELISA, Western Blot, or flow cytometry.
  • point-of-care systems such as microfluidic devices or lateral flow assay are used to the detect and measure the biomarkers.
  • other techniques can be used to detect and measure biomarker concentrations.
  • a concentration of CRP within a range of 160,000 ng/ml to 8,000,000 ng/ml is indicative of bacterial infection.
  • a concentration of neutrophil elastase within a range of 23,000 to 720,000 ng/ml is indicative of bacterial infection.
  • a concentration of p-Defensin 2 within a range of 7,600 to 30,000 ng/ml is indicative of bacterial infection.
  • a concentration of IL-8 within a range of 35,000 to 800,000 ng/ml is indicative of bacterial infection.
  • a concentration of MPO within a range of 20,000,000 to 200,000,000 ng/ml is indicative of bacterial infection.
  • a concentration of TREM-2 within a range of 3,000 to 360,000 ng/ml is indicative of bacterial infection.
  • a concentration of G-CSF within a range of 4,300 to 50,000 ng/ml is indicative of bacterial infection.
  • a concentration of SAA within a range of 45,000 to 300,000 ng/ml is indicative of bacterial infection.
  • concentration of IP-10 within a range of 50 to 800 ng/ml is indicative of viral infection.
  • concentration of MxA within a range of 0.6 to 320 ng/ml is indicative of viral infection.
  • a concentration of TRAIL within a range of 3,000 to 22,000 ng/ml is indicative of viral infection.
  • two or more biomarkers in combination are tested to indicate the cause as either a bacterial infection or viral infection.
  • the combination of I P-10 and CRP has 100% sensitivity and 93% specificity.
  • the present invention relates to a method for establishing reference diagnostic values to assist with sinonasal inflammatory profiling.
  • the method involves: a) collecting biological samples from a cohort of patients; b) detecting and measuring the concentration of biomarkers in the biological samples that are associated with viral infections and bacterial infections; and c) computing reference diagnostic values based on the measured concentrations.
  • the cohort preferably comprises healthy (control) patients, patients with viral infection, patients with bacterial infections.
  • the cohort comprises patients with mixed infections, however, it is to be appreciated that cohorts without mixed infections can be preferable.
  • the biological samples are sinonasal samples including sinonasal secretions, sinonasal aspirates and sinonasal tissue cultures.
  • the sinonasal sample is collected by forced exhalation or absorptive strips.
  • the dilution ratio of the sinonasal sample is a standardised dilution ratio for accurate testing.
  • the biomarkers measured are preferably selected from a biomarker panel comprising Human p-Defensin 2 (BD-2), C-reactive protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte- Colony Stimulating Factor (G-CSF), Granzyme B, lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Matrix metalloproteinase-1 (MMP-1 ), Matrix metalloproteinase-8 (MMP-8), Myeloperoxidase (MPO), Human Myxovirus Resistance protein 1 (MxA), Neutrophil Elastase (NE), Serum Amyloid A (SAA), Tumor necrosis factor A (TNFa), Tumor necrosis factor-related apoptosis
  • the invention relates to a kit for detecting biomarkers associated with sinonasal inflammatory conditions caused by viral and bacterial infections.
  • the kit comprises one or more agents for detecting protein biomarkers associated with viral and bacterial infections wherein one or more protein biomarkers are selected from Human p-Defensin 2 (BD-2), C-reactive protein (CRP), Eotaxin-2, Eotaxin-3, Granulocyte- Colony Stimulating Factor (G-CSF), Granzyme B, lnterleukin-3 (IL-3), lnterleukin-5 (IL-5), lnterleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-37 (IL-37), Interferon gamma inducible protein-10 (IP-10), Matrix metalloproteinase- 1 (MMP-1 ), Matrix metalloproteinase-8 (MMP-8), Myeloperoxidase (MPO), Human
  • BD-2 Human p-Def
  • the kit comprises one or more agents for detecting nucleic acid biomarkers associated with viral and bacterial infections.
  • the one or more agents are appropriate for detecting RNA using hybridisation techniques and microarrays.
  • PCR Polymerase Chain Reaction
  • Sinonasal samples were collected from patients diagnosed with sinonasal inflammatory conditions through forced exhalation of nasal secretions and from healthy patients using absorptive strips. Collected sinonasal samples were stored at about minus 80°C under appropriate conditions prior to biomarker quantification.
  • the concentrations of the protein biomarkers including BD-2, CRP, Eotaxin-2, Eotaxin-3, G-CSF, Granzyme B, IL-3, IL-5, IL-8, IL-10, IL-37, IP-10, MMP-1 , MMP-8, MPO, MxA, NE, SAA, TNFa, TRAIL, TREM-1 , and TREM-
  • a sandwich enzyme immunoassay kit by BioVendor (Lot: E21 -049) was used to detect and quantify the concentration of Human MxA protein in the collected sinonasal samples. Reagents were brought to room temperature prior to use. The master standard was reconstituted with dilution buffer and gently agitated for 15 min. The master standard was aliquoted in duplicates into a 96-well microtiter plate at concentrations of 6 ng/ml, 3 ng/ml, 1 .5 ng/ml, 0.75 ng/ml and 0.375 ng/ml and at a final volume of 100 ml.
  • Sinonasal samples collected from each patient group were aliquoted in duplicates into the microtiter plate at different dilutions.
  • the microtiter plate was incubated at room temperature for 1 hour at 300 RPM.
  • the wells were washed 3 times with the wash solution and the plate was inverted and tapped against a paper towel to remove excess solution.
  • One hundred microlitres of Biotin Labelled Anti-MxA Antibody solution was pipetted into each well and the plate was incubated at room temperature for 1 hour at 300 RPM.
  • the wells were washed again 3 times with 350 ml of the wash solution, inverted and tapped against a paper towel to remove excess solution.
  • Streptavidin-HRP conjugate was pipetted into each well and the plate was incubated for 1 hour at room temperature at 300 RPM. The wells were washed 5 times with 350 ml of wash solution and 100 ml of substrate solution was pipetted into each well. The plate was incubated for 20 minutes at room temperature. One hundred microlitres of stop solution was pipetted into each well to stop the enzymatic reaction and optical density was measured at 450/630 nm.
  • a sandwich enzyme immunoassay kit by Alpha Diagnostic (Cat: 100-250-BD2) was used to detect and quantify the concentration of p-Defensin 2 protein in the collected sinonasal samples. Reagents were brought to room temperature prior to use and reconstituted and diluted as per manufacturer’s instructions. One hundred microlitres of each collected sample, controls and standards were aliquoted in duplicates into a microtiter plate and incubated for 60 min at room temperature. The wells were washed 4 times with wash solution and excess liquid was decanted onto a paper towel after the final wash. One hundred microlitres of Anti-BD-2 Detecting Antibody was pipetted into each well and the plate was incubated at room temperature for 60 min.
  • a sandwich enzyme immunoassay kit by Hycult Biotech (Lot: 27909K0221 -K) was used to detect and quantify the concentration of Neutrophil Elastase protein in the collected sinonasal samples. Reagents were brought to room temperature prior to use and reconstituted and diluted as per manufacturer’s instructions. One hundred microlitres of each collected sample, controls and standards were aliquoted in duplicates into a microtiter plate and incubated for 1 hour at room temperature. The wells were washed 4 times with wash buffer and excess liquid was decanted onto a paper towel after the final wash. Two hundred microlitres of wash buffer was pipetted into each well and the plate was incubated for 20 sec at room temperature.
  • the ROC curve is used to generate three values of importance: Min corresponding to lowest sensitivity and maximum specificity, Max corresponding to highest sensitivity and lowest specificity, and Peak corresponding to balance sensitivity and specificity.
  • a Min cutoff, Max cutoff, and Peak cutoff values are determined corresponding to the biomarker concentration at Min, Max, and Peak, respectively.
  • ROC curves (Fig 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22) were generated and reference diagnostic range thresholds including cut off values, associated sensitivity and specificity values, accuracy, area under the curve (AUC), 95% confidence interval and P-value (Fig 23) were determined from the ROC curves.
  • Combinations of two biomarkers were also tested to indicate the cause as either a bacterial infection or viral infection and the reference diagnostic range thresholds are shown in Figure 24.
  • the combination of IP-10 and CRP has 100% sensitivity and 93% specificity at the specified cutoff concentrations.
  • ROC curves (Fig 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62) were generated and reference diagnostic range thresholds including cut off values, associated sensitivity and specificity values, area under the curve (AUC), 95% confidence interval and P-value (Fig 63) were determined from the ROC curves. Combinations of two biomarkers were also tested to indicate the cause as either a bacterial infection or viral infection and the reference diagnostic range thresholds are shown in Figure 64.
  • ROC curves (Fig 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86) were generated and reference diagnostic range thresholds including cut off values, associated sensitivity and specificity values, area under the curve (AUC), 95% confidence interval and P-value (Fig 87) were determined from the ROC curves. Combinations of two biomarkers were also tested to indicate the cause as either a bacterial infection or viral infection and the reference diagnostic range thresholds are shown in Figure 88.
  • the results of Study 1 demonstrate that SAA and TREM-2 are suitable as bacterial biomarkers
  • the results of Study 2 demonstrate that SAA and TREM-2 are suitable as viral biomarkers. Accordingly, the results of the Studies support embodiments in which SAA and TREM-2 are bacterial biomarkers only and support embodiments in which SAA and TREM-2 are viral biomarkers only or embodiments where SAA and TREM-2 are bacterial or viral biomarkers.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Analytical Chemistry (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • General Health & Medical Sciences (AREA)
  • Food Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Cell Biology (AREA)
  • Medicinal Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Zoology (AREA)
  • Virology (AREA)
  • Wood Science & Technology (AREA)
  • Biophysics (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

La présente invention concerne une méthode destinée au diagnostic d'un état inflammatoire rhinosinusien chez un patient provoqué par une infection virale ou bactérienne. Le procédé comprend la mesure de concentrations d'une combinaison de biomarqueurs dans un échantillon rhinosinusien recueilli sur un patient et la comparaison de la concentration des biomarqueurs à une ou plusieurs valeurs de diagnostic de référence. L'invention concerne également un procédé pour établir des valeurs de diagnostic de référence, une méthode pour déterminer un traitement approprié et des kits pour le diagnostic d'états inflammatoires rhinosinusiens provoqués par des infections virales et/ou bactériennes.
PCT/AU2023/051279 2022-12-08 2023-12-08 Méthodes destinées au diagnostic et au traitement d'une infection rhinosinusienne virale et bactérienne WO2024119245A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202263431124P 2022-12-08 2022-12-08
US63/431,124 2022-12-08

Publications (1)

Publication Number Publication Date
WO2024119245A1 true WO2024119245A1 (fr) 2024-06-13

Family

ID=91378269

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/AU2023/051279 WO2024119245A1 (fr) 2022-12-08 2023-12-08 Méthodes destinées au diagnostic et au traitement d'une infection rhinosinusienne virale et bactérienne

Country Status (1)

Country Link
WO (1) WO2024119245A1 (fr)

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014137858A1 (fr) * 2013-03-07 2014-09-12 Rapid Pathogen Screening, Inc. Procédé et dispositif de détection combinée d'infections virales et bactériennes
WO2016092554A1 (fr) * 2014-12-11 2016-06-16 Memed Diagnostics Ltd. Combinaisons de marqueurs de diagnostic d'infections et leurs méthodes d'utilisation
WO2018029690A1 (fr) * 2016-08-10 2018-02-15 Memed Diagnostics Ltd. Système et procédé d'analyse de données biologiques
US20210318311A1 (en) * 2020-04-13 2021-10-14 Mbio Diagnostics, Inc. Simultaneous detection of humoral and inflammatory biomarkers
WO2021219670A1 (fr) * 2020-04-27 2021-11-04 Abacuslabs Ltd. Méthode de distinction d'individus sains d'individus présentant des états infectieux ou inflammatoires
JP2022061491A (ja) * 2020-10-06 2022-04-18 国立大学法人福井大学 慢性副鼻腔炎の患者を分類するためのデータの取得方法、およびその利用
WO2022184942A2 (fr) * 2021-03-05 2022-09-09 Oncimmune Germany Gmbh Biomarqueurs

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014137858A1 (fr) * 2013-03-07 2014-09-12 Rapid Pathogen Screening, Inc. Procédé et dispositif de détection combinée d'infections virales et bactériennes
WO2016092554A1 (fr) * 2014-12-11 2016-06-16 Memed Diagnostics Ltd. Combinaisons de marqueurs de diagnostic d'infections et leurs méthodes d'utilisation
WO2018029690A1 (fr) * 2016-08-10 2018-02-15 Memed Diagnostics Ltd. Système et procédé d'analyse de données biologiques
US20210318311A1 (en) * 2020-04-13 2021-10-14 Mbio Diagnostics, Inc. Simultaneous detection of humoral and inflammatory biomarkers
WO2021219670A1 (fr) * 2020-04-27 2021-11-04 Abacuslabs Ltd. Méthode de distinction d'individus sains d'individus présentant des états infectieux ou inflammatoires
JP2022061491A (ja) * 2020-10-06 2022-04-18 国立大学法人福井大学 慢性副鼻腔炎の患者を分類するためのデータの取得方法、およびその利用
WO2022184942A2 (fr) * 2021-03-05 2022-09-09 Oncimmune Germany Gmbh Biomarqueurs

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
ANONYMOUS: "GeneChip™ WT Pico Reagent Kit - USER GUIDE", APPLIEDBIOSYSTEMS, 26 October 2020 (2020-10-26), XP093182004, Retrieved from the Internet <URL:https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017893_703262_GeneChipWT_Pico_UG.pdf> *
MUEHLING LYNDSEY M., HEYMANN PETER W., CARPER HOLLIDAY, MURPHY DEBORAH D., RAJADHYAKSHA EVAN, KENNEDY JOSHUA, EARLY STEPHEN V., SO: "Cluster analysis of nasal cytokines during rhinovirus infection identifies different immunophenotypes in both children and adults with allergic asthma", CLINICAL & EXPERIMENTAL ALLERGY, WILEY INTERSCIENCE, UK, vol. 52, no. 10, 1 October 2022 (2022-10-01), UK , pages 1169 - 1182, XP093182002, ISSN: 0954-7894, DOI: 10.1111/cea.14176 *

Similar Documents

Publication Publication Date Title
Endo et al. Usefulness of presepsin in the diagnosis of sepsis in a multicenter prospective study
Kim et al. Plasma neutrophil gelatinase-associated lipocalin: a marker of acute pyelonephritis in children
Zehnder et al. A critical analysis of research methods to study clinical molecular biomarkers in Endodontic research
Eribo et al. Host urine immunological biomarkers as potential candidates for the diagnosis of tuberculosis
Short et al. Optimising the collection of female genital tract fluid for cytokine analysis in pregnant women
CN111721941A (zh) 一种用于判断脓毒症感染情况的装置及其应用
Jaumdally et al. Lower genital tract cytokine profiles in South African women living with HIV: influence of mucosal sampling
JP2018072163A (ja) 間質性膀胱炎鑑別用マーカー及びその用途
Kumar et al. Dynamic alteration in the vaginal secretory proteome across the early and mid-trimesters of pregnancy
WO2024183386A1 (fr) Réactif de détection à base de chaîne d&#39;oligosaccharide pour détecter un cancer de la prostate, procédé de préparation et utilisation
Alves et al. Evaluation of potential biomarkers for the diagnosis and monitoring of systemic lupus erythematosus using the cytometric beads array (CBA)
CN113186311A (zh) 阴道微生物在慢性盆腔痛综合征鉴别诊断中的应用
WO2024119245A1 (fr) Méthodes destinées au diagnostic et au traitement d&#39;une infection rhinosinusienne virale et bactérienne
CN114636826B (zh) Cd177+中性粒细胞在制备新生儿坏死性小肠结肠炎检测产品中的应用
KR20200049647A (ko) 반려동물에서 종양질병의 보조적 진단을 위한 바이오마커 검사방법
Casini et al. Use of transcriptomics for diagnosis of infections and sepsis in children: A narrative review
RU2684287C1 (ru) Способ прогнозирования риска развития псориаза 2 типа у женщин в постменопаузе
Bullarbo et al. Self-diagnosis of vulvovaginal candidiasis is poor—a comparison of diagnostic methods introducing β-glucan as a complement
CN110373466B (zh) 一种标记物组合及其在制备结直肠癌诊断试剂中的应用
KR20210026852A (ko) 비용종을 갖는 비부비동염의 진단을 위한 정보 제공 방법
CN113252910B (zh) Timp3在焦虑抑郁症诊断中的应用
EP3063542B1 (fr) Méthode de diagnostic d&#39;une affection inflammatoire touchant l&#39;appareil génital féminin
Ali et al. Gingival crevicular fluid volume and protein concentration: A biomarker tool for predicting periodontal diseases progression and severity
Zaheer et al. Potential for and challenges of menstrual blood as a non-invasive diagnostic specimen: Current status and future directions
Salvagno et al. The role of acute phase proteins for predicting SARS-CoV-2 positivity upon emergency department admission

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23899143

Country of ref document: EP

Kind code of ref document: A1

DPE1 Request for preliminary examination filed after expiration of 19th month from priority date (pct application filed from 20040101)