Development and Validation of Indirect Enzyme-Linked Immunosorbent Assays for Detecting Antibodies to SARS-CoV-2 in Cattle, Swine, and Chicken

Abhinay Gontu; Erika A Marlin; Santhamani Ramasamy; Sabarinath Neerukonda; Gayatri Anil; Jasmine Morgan; Meysoon Quraishi; Chen Chen; Veda Sheersh Boorla; Ruth H Nissly; Padmaja Jakka; Shubhada K Chothe; Abirami Ravichandran; Nishitha Kodali; Saranya Amirthalingam; Lindsey LaBella; Kathleen Kelly; Pazhanivel Natesan; Allen M Minns; Randall M Rossi; Jacob R Werner; Ernest Hovingh; Scott E Lindner; Deepanker Tewari; Vivek Kapur; Kurt J Vandegrift; Costas D Maranas; Meera Surendran Nair; Suresh V Kuchipudi

doi:10.3390/v14071358

Development and Validation of Indirect Enzyme-Linked Immunosorbent Assays for Detecting Antibodies to SARS-CoV-2 in Cattle, Swine, and Chicken

Viruses. 2022 Jun 22;14(7):1358. doi: 10.3390/v14071358.

Authors

Abhinay Gontu^{1

2}, Erika A Marlin^{1

3}, Santhamani Ramasamy¹, Sabarinath Neerukonda⁴, Gayatri Anil¹, Jasmine Morgan¹, Meysoon Quraishi¹, Chen Chen⁵, Veda Sheersh Boorla⁵, Ruth H Nissly², Padmaja Jakka^{1

2}, Shubhada K Chothe¹, Abirami Ravichandran⁶, Nishitha Kodali^{1

7}, Saranya Amirthalingam^{1

7}, Lindsey LaBella¹, Kathleen Kelly², Pazhanivel Natesan⁸, Allen M Minns^{7

9}, Randall M Rossi⁷, Jacob R Werner¹⁰, Ernest Hovingh¹, Scott E Lindner^{7

9}, Deepanker Tewari¹¹, Vivek Kapur^{7

10

12}, Kurt J Vandegrift^{7

12

13}, Costas D Maranas⁵, Meera Surendran Nair^{1

2}, Suresh V Kuchipudi^{1

2

7

12}

Affiliations

¹ Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, PA 16802, USA.
² Animal Diagnostic Laboratory, Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, PA 16802, USA.
³ Clinical & Diagnostic Assay Development Group, Pfizer, Pearl River, NY 10965, USA.
⁴ U.S. Department of Health and Human Services, Silver Spring, MD 20993, USA.
⁵ Department of Chemical Engineering, The Pennsylvania State University, University Park, PA 16802, USA.
⁶ Department of Integrative and Biomedical Physiology, The Pennsylvania State University, University Park, PA 16802, USA.
⁷ Huck Institute of Life Sciences, The Pennsylvania State University, University Park, PA 16802, USA.
⁸ Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Chennai 600007, India.
⁹ Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA.
¹⁰ Department of Animal Science, The Pennsylvania State University, University Park, PA 16802, USA.
¹¹ Pennsylvania Department of Agriculture, Pennsylvania Veterinary Laboratory, Harrisburg, PA 17110, USA.
¹² Center for Infectious Disease Dynamics, The Pennsylvania State University, University Park, PA 16802, USA.
¹³ Department of Biology, The Pennsylvania State University, University Park, PA 16802, USA.

Abstract

Multiple domestic and wild animal species are susceptible to SARS-CoV-2 infection. Cattle and swine are susceptible to experimental SARS-CoV-2 infection. The unchecked transmission of SARS-CoV-2 in animal hosts could lead to virus adaptation and the emergence of novel variants. In addition, the spillover and subsequent adaptation of SARS-CoV-2 in livestock could significantly impact food security as well as animal and public health. Therefore, it is essential to monitor livestock species for SARS-CoV-2 spillover. We developed and optimized species-specific indirect ELISAs (iELISAs) to detect anti-SARS-CoV-2 antibodies in cattle, swine, and chickens using the spike protein receptor-binding domain (RBD) antigen. Serum samples collected prior to the COVID-19 pandemic were used to determine the cut-off threshold. RBD hyperimmunized sera from cattle (n = 3), swine (n = 6), and chicken (n = 3) were used as the positive controls. The iELISAs were evaluated compared to a live virus neutralization test using cattle (n = 150), swine (n = 150), and chicken (n = 150) serum samples collected during the COVID-19 pandemic. The iELISAs for cattle, swine, and chicken were found to have 100% sensitivity and specificity. These tools facilitate the surveillance that is necessary to quickly identify spillovers into the three most important agricultural species worldwide.

Keywords: ELISA; SARS-CoV-2; cattle; chicken; serology; surveillance; swine.

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antibodies, Viral
COVID-19* / diagnosis
COVID-19* / veterinary
Cattle
Chickens
Enzyme-Linked Immunosorbent Assay
Humans
Pandemics / prevention & control
SARS-CoV-2*
Spike Glycoprotein, Coronavirus
Swine

Substances

Antibodies, Viral
Spike Glycoprotein, Coronavirus
spike protein, SARS-CoV-2

Abstract

Publication types

MeSH terms

Substances

Grants and funding