Developing and validating SARS‐CoV‐2 assays for nonhuman primate surveillance
Introduction In early 2020, the California National Primate Research Center implemented surveillance to address the threat of SARS‐CoV‐2 infection in its nonhuman primate colony. Materials/Methods To detect antiviral antibodies, multi‐antigen assays were developed and validated on enzyme immunoassay...
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| Published in: | Journal of medical primatology Vol. 51; no. 5; pp. 264 - 269 |
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| Main Authors: | , , , , , |
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| Language: | English |
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| ISSN: | 0047-2565, 1600-0684, 1600-0684 |
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| Abstract | Introduction
In early 2020, the California National Primate Research Center implemented surveillance to address the threat of SARS‐CoV‐2 infection in its nonhuman primate colony.
Materials/Methods
To detect antiviral antibodies, multi‐antigen assays were developed and validated on enzyme immunoassay and multiplex microbead immunofluorescent assay (MMIA) platforms. To detect viral RNA, RT‐PCR was also performed.
Results/Conclusion
Using a 4plex, antibody was identified in 16/16 experimentally infected animals; and specificity for spike, nucleocapsid, receptor binding domain, and whole virus antigens was 95.2%, 93.8%, 94.3%, and 97.1%, respectively on surveillance samples. Six laboratories compared this MMIA favorably with nine additional laboratory‐developed or commercially available assays. Using a screen and confirm algorithm, 141 of the last 2441 surveillance samples were screen‐reactive requiring confirmatory testing. Although 35 samples were reactive to either nucleocapsid or spike; none were reactive to both. Over 20 000 animals have been tested and no spontaneous infections have so far been confirmed across the NIH sponsored National Primate Research Centers. |
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| AbstractList | IntroductionIn early 2020, the California National Primate Research Center implemented surveillance to address the threat of SARS‐CoV‐2 infection in its nonhuman primate colony.Materials/MethodsTo detect antiviral antibodies, multi‐antigen assays were developed and validated on enzyme immunoassay and multiplex microbead immunofluorescent assay (MMIA) platforms. To detect viral RNA, RT‐PCR was also performed.Results/ConclusionUsing a 4plex, antibody was identified in 16/16 experimentally infected animals; and specificity for spike, nucleocapsid, receptor binding domain, and whole virus antigens was 95.2%, 93.8%, 94.3%, and 97.1%, respectively on surveillance samples. Six laboratories compared this MMIA favorably with nine additional laboratory‐developed or commercially available assays. Using a screen and confirm algorithm, 141 of the last 2441 surveillance samples were screen‐reactive requiring confirmatory testing. Although 35 samples were reactive to either nucleocapsid or spike; none were reactive to both. Over 20 000 animals have been tested and no spontaneous infections have so far been confirmed across the NIH sponsored National Primate Research Centers. In early 2020, the California National Primate Research Center implemented surveillance to address the threat of SARS-CoV-2 infection in its nonhuman primate colony. To detect antiviral antibodies, multi-antigen assays were developed and validated on enzyme immunoassay and multiplex microbead immunofluorescent assay (MMIA) platforms. To detect viral RNA, RT-PCR was also performed. Using a 4plex, antibody was identified in 16/16 experimentally infected animals; and specificity for spike, nucleocapsid, receptor binding domain, and whole virus antigens was 95.2%, 93.8%, 94.3%, and 97.1%, respectively on surveillance samples. Six laboratories compared this MMIA favorably with nine additional laboratory-developed or commercially available assays. Using a screen and confirm algorithm, 141 of the last 2441 surveillance samples were screen-reactive requiring confirmatory testing. Although 35 samples were reactive to either nucleocapsid or spike; none were reactive to both. Over 20 000 animals have been tested and no spontaneous infections have so far been confirmed across the NIH sponsored National Primate Research Centers. Introduction In early 2020, the California National Primate Research Center implemented surveillance to address the threat of SARS‐CoV‐2 infection in its nonhuman primate colony. Materials/Methods To detect antiviral antibodies, multi‐antigen assays were developed and validated on enzyme immunoassay and multiplex microbead immunofluorescent assay (MMIA) platforms. To detect viral RNA, RT‐PCR was also performed. Results/Conclusion Using a 4plex, antibody was identified in 16/16 experimentally infected animals; and specificity for spike, nucleocapsid, receptor binding domain, and whole virus antigens was 95.2%, 93.8%, 94.3%, and 97.1%, respectively on surveillance samples. Six laboratories compared this MMIA favorably with nine additional laboratory‐developed or commercially available assays. Using a screen and confirm algorithm, 141 of the last 2441 surveillance samples were screen‐reactive requiring confirmatory testing. Although 35 samples were reactive to either nucleocapsid or spike; none were reactive to both. Over 20 000 animals have been tested and no spontaneous infections have so far been confirmed across the NIH sponsored National Primate Research Centers. In early 2020, the California National Primate Research Center implemented surveillance to address the threat of SARS-CoV-2 infection in its nonhuman primate colony.INTRODUCTIONIn early 2020, the California National Primate Research Center implemented surveillance to address the threat of SARS-CoV-2 infection in its nonhuman primate colony.To detect antiviral antibodies, multi-antigen assays were developed and validated on enzyme immunoassay and multiplex microbead immunofluorescent assay (MMIA) platforms. To detect viral RNA, RT-PCR was also performed.MATERIALS/METHODSTo detect antiviral antibodies, multi-antigen assays were developed and validated on enzyme immunoassay and multiplex microbead immunofluorescent assay (MMIA) platforms. To detect viral RNA, RT-PCR was also performed.Using a 4plex, antibody was identified in 16/16 experimentally infected animals; and specificity for spike, nucleocapsid, receptor binding domain, and whole virus antigens was 95.2%, 93.8%, 94.3%, and 97.1%, respectively on surveillance samples. Six laboratories compared this MMIA favorably with nine additional laboratory-developed or commercially available assays. Using a screen and confirm algorithm, 141 of the last 2441 surveillance samples were screen-reactive requiring confirmatory testing. Although 35 samples were reactive to either nucleocapsid or spike; none were reactive to both. Over 20 000 animals have been tested and no spontaneous infections have so far been confirmed across the NIH sponsored National Primate Research Centers.RESULTS/CONCLUSIONUsing a 4plex, antibody was identified in 16/16 experimentally infected animals; and specificity for spike, nucleocapsid, receptor binding domain, and whole virus antigens was 95.2%, 93.8%, 94.3%, and 97.1%, respectively on surveillance samples. Six laboratories compared this MMIA favorably with nine additional laboratory-developed or commercially available assays. Using a screen and confirm algorithm, 141 of the last 2441 surveillance samples were screen-reactive requiring confirmatory testing. Although 35 samples were reactive to either nucleocapsid or spike; none were reactive to both. Over 20 000 animals have been tested and no spontaneous infections have so far been confirmed across the NIH sponsored National Primate Research Centers. |
| Author | Carpenter, Amanda Roberts, Jeffrey Nham, Peter Halley, Bryson Yee, JoAnn Van Rompay, Koen K. A. |
| AuthorAffiliation | 2 Department of Pathology, Microbiology & Immunology, School of Veterinary Medicine University of California, Davis Davis California USA 1 Primate Assay Laboratory, California National Primate Research Center University of California, Davis Davis California USA 3 Department Medicine and Epidemiology, School of Veterinary Medicine University of California, Davis Davis California USA |
| AuthorAffiliation_xml | – name: 2 Department of Pathology, Microbiology & Immunology, School of Veterinary Medicine University of California, Davis Davis California USA – name: 3 Department Medicine and Epidemiology, School of Veterinary Medicine University of California, Davis Davis California USA – name: 1 Primate Assay Laboratory, California National Primate Research Center University of California, Davis Davis California USA |
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| Snippet | Introduction
In early 2020, the California National Primate Research Center implemented surveillance to address the threat of SARS‐CoV‐2 infection in its... In early 2020, the California National Primate Research Center implemented surveillance to address the threat of SARS-CoV-2 infection in its nonhuman primate... IntroductionIn early 2020, the California National Primate Research Center implemented surveillance to address the threat of SARS‐CoV‐2 infection in its... |
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| SubjectTerms | Animals Antibodies, Viral antibody Antigens Antiviral drugs COVID-19 - diagnosis diagnostic testing algorithm Enzyme immunoassay Laboratories Microspheres Nucleocapsids Original RNA, Viral RT‐PCR SARS-CoV-2 Sensitivity and Specificity Severe acute respiratory syndrome coronavirus 2 Surveillance |
| Title | Developing and validating SARS‐CoV‐2 assays for nonhuman primate surveillance |
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