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US-20260126442-A1 - METHODS FOR MONITORING IMMUNE STATUS OF A POPULATION

US20260126442A1US 20260126442 A1US20260126442 A1US 20260126442A1US-20260126442-A1

Abstract

The present invention relates to methods for monitoring the immune status of a population by detecting and/or quantifying antibodies present in wastewater. The invention further provides methods and kits for monitoring the presence of an antibody to SARS-CoV-2 or a variant thereof within a population by detecting and/or quantifying SARS-CoV-2-specific antibodies in wastewater.

Inventors

  • Cynthia GIBAS
  • Jessica SCHLUETER
  • William Taylor
  • Marie LittleFawn AGAN

Assignees

  • THE UNIVERSITY OF NORTH CAROLINA AT CHARLOTTE

Dates

Publication Date
20260507
Application Date
20231024

Claims (20)

  1. 1 . A method of detecting the presence of an antibody to an antigen of interest within a population comprising (a) contacting a wastewater sample from a population with an antigen of interest, and (b) detecting binding of the antibody to the antigen of interest, thereby detecting the antibody to the antigen of interest within the population.
  2. 2 . The method of claim 1 , wherein the antigen of interest is from a pathogen, optionally a bacterium, parasite, fungus, or virus.
  3. 3 . (canceled)
  4. 4 . The method of claim 3 , wherein the virus is a coronavirus, optionally SARS-CoV-2 or variant thereof, optionally wherein the antigen of interest is the spike protein and/or nucleocapsid protein.
  5. 5 - 6 . (canceled)
  6. 7 . The method of claim 1 , wherein the antigen of interest is from a vaccine.
  7. 8 . The method of claim 1 , wherein the step of detecting binding of the antibody to the antigen of interest comprises using an immunodiffusion assay, immunoelectrophoretic assay, light scattering immunoassay, agglutination assay, or labeled immunoassay, optionally wherein the labeled immunoassay is an enzyme-linked immunosorbent assay.
  8. 9 . (canceled)
  9. 10 . The method of claim 1 , further comprising a prestep of concentrating the wastewater sample from the population, optionally wherein the step of concentrating the wastewater sample comprises ultrafiltration of the wastewater sample.
  10. 11 . (canceled)
  11. 12 . The method of claim 1 , wherein the wastewater sample is fresh or frozen and/or wherein the antibody is an IgA and/or IgG antibody.
  12. 13 . (canceled)
  13. 14 . The method of claim 1 , further comprising (c) repeating (a)-(b) 2 or more times to monitor the presence of the antibody to the antigen of interest within the population.
  14. 15 . The method of claim 1 , further comprising quantifying the amount of antibody in the wastewater sample.
  15. 16 . The method of claim 2 , further comprising detecting and/or quantifying the amount of the pathogen present in the wastewater sample.
  16. 17 . A method of determining immune status of a population to a pathogen comprising (a) contacting a wastewater sample from a population with at least one antigen from the pathogen; and (b) detecting binding of antibodies present in the wastewater sample to the at least one antigen from the pathogen, thereby determining the immune status of the population to the pathogen.
  17. 18 . The method of claim 17 , wherein the pathogen is a bacterium, parasite, fungus, or virus.
  18. 19 . The method of claim 18 , wherein the virus is a coronavirus, optionally wherein the coronavirus is SARS-CoV-2 or variant thereof, optionally wherein the at least one antigen is the spike protein and/or nucleocapsid protein.
  19. 20 - 21 . (canceled)
  20. 22 . The method of claim 17 , wherein the at least one antigen is from a vaccine.

Description

STATEMENT OF PRIORITY The present invention claims the benefit, under 35 U.S.C. § 119 (e), of U.S. Provisional Application No. 63/419,205, filed Oct. 25, 2022, the entire contents of which are incorporated by reference herein. STATEMENT REGARDING ELECTRONIC FILING OF A SEQUENCE LISTING A Sequence Listing in XML format, entitled 9812-11WO_ST26.xml, 29,476 bytes in size, generated on Oct. 21, 2023 and filed herewith, is hereby incorporated by reference in its entirety for its disclosures. FIELD OF THE INVENTION The present invention relates to the field of wastewater surveillance for monitoring immune status and infectious disease in populations. BACKGROUND OF THE INVENTION Seroprevalence surveillance plays a critical role in detection and epidemiology of infectious disease outbreaks (Choisy, et al. (2019) Open Forum Infect. Dis. 6: ofz030; Wilson et al. (2012) CMAJ 184: E70-6). Serological evidence of infection can also be used as an indicator of likelihood of zoonotic spillover and precursor to emergence of novel diseases (Li et al. (2019) Biosaf. Health 1:84-90). The onset of the COVID-19 pandemic led to a call for a Global Immunological Observatory (Mina et al. (2020) Elife 9: e58989) that would provide information about susceptibility of populations based on aggregate serological data, allowing for public health action in advance of waves of infection. However, there are several requirements which are barriers to large-scale data collection and the achievement of such an aim. One of these is the need to collect large numbers of serological samples from individual patients, which implies the need for patient participation, or to reliably obtain material from clinical discards. Such requirements are especially challenging in low and middle income countries (Haselbeck et al. (2022) Pathogens 11 (7) 732) but can pose a challenge even in resource-rich areas. In the case of the SARS-CoV-2 virus, a massive amount of genomic surveillance data was collected as governments required testing and made free testing available, but as pandemic mitigation measures have declined or ended, clinical test discards for sequencing have become difficult to obtain and surveillance by genomic sequencing has declined significantly. In turn, health departments have turned to sequencing and viral variant identification out of wastewater as a non-invasive method that can be conducted independent of patient choices about treatment and testing. Wastewater surveillance has proven useful at both the regional scale for detection of SARS-CoV-2 infection trends (Peccia et al. (2020) Nat. Biotechnol. 38:1164-1167; Wu et al. (2022) Sci. Total Environ. 805:150121; Weidhaas et al. (2021) Sci. Total Environ. 775:145790), and at the local scale for guiding rapid response to emerging outbreaks (Gibas et al. (2021) Sci. Total Environ. 782:146749; Johnson et al. (2022) FEMS Microbes 24:3:xtac024). The main modes of wastewater surveillance during the COVID-19 pandemic have been quantitative detection and variant sequencing of viral RNA isolated from wastewater. However, the potential for monitoring other immunologically relevant molecules in wastewater has not been as extensively explored. Immunoassay systems have been widely used as a biotechnology tool for detection of SARS-CoV-2 RNA (Fourati et al. (2022) J. Clin. Virol. 146:105048; Drain et al. (2021) Infect. Dis. Ther. 10:753-61) and virus in clinical samples. Samples for immunoassays are typically collected from serum (Guerrero-Esteban et al. (2022) Talanta 247:123543), but there have also been recent advances in use of immunosensors targeting the viral antigen protein in wastewater (Thongpradit et al. (2022) Int. J. Environ. Res. Public Health 19 (13): 7783; Lu et al. (2021) Sci. Total Environ. 77:146239). Antibody measurements from fecal samples have been described as a non-invasive means to rapidly monitor the immune status of wildlife (Watt et al. (2016) Ecol. Evol. 6:56-67). Similarly, measurements from fecal samples are being used for studying various aspects of human health (Frehn et al. (2014) PLoS ONE 9: e106750; Lin et al. (2018) J. Transl. Med. 16:359), as fecal antibody levels are correlated with antibody levels in serum (Kolmannskog & Haneberg (1985) Int. Arch. Allergy Appl. Immunol. 76:133-137), and have been demonstrated as a potential biomarker for detection of viral (Niedrig et al. (2018) BMC Infect. Dis. 18:707) and parasitic infections (Nagaoka et al. (2021) Vaccines (Basel) 9 (7): 778) in individual human urine samples. Infection by SARS-CoV-2 elicits the secretion of mucosal IgG and IgA antibodies (Lehmann et al. (2021) Mucosal Immunol. 14:1381-1392). The IgA response waxes and wanes rapidly, being detectable in serum within days of infection, before falling below the limit of detection within weeks. In contrast, the signal of the IgG response in serum rises more slowly and may persist for months (Townsend et al. (2021) Lancet Microbe 2: e666-e675), potentially remaining ab