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Development of an Indirect ELISA for the Detection of SARS-CoV-2 Antibodies in Cats.
Bold, Dashzeveg; Roman-Sosa, Gleyder; Gaudreault, Natasha N; Zayat, Batsukh; Pogranichniy, Roman M; Richt, Juergen A.
  • Bold D; Department of Diagnostic Medicine/Pathobiology, Kansas State University, Manhattan, KS, United States.
  • Roman-Sosa G; Department of Diagnostic Medicine/Pathobiology, Kansas State University, Manhattan, KS, United States.
  • Gaudreault NN; Institut für Virologie, Fachbereich Veterinärmedizin, Justus-Liebig-Universität Giessen, Giessen, Germany.
  • Zayat B; Department of Diagnostic Medicine/Pathobiology, Kansas State University, Manhattan, KS, United States.
  • Pogranichniy RM; Institute of Veterinary Medicine, Mongolian University of Life Sciences, Ulaanbaatar, Mongolia.
  • Richt JA; Veterinary Diagnostic Laboratory, Department of Diagnostic Medicine/Pathobiology, Kansas State University, Manhattan, KS, United States.
Front Vet Sci ; 9: 864884, 2022.
Article in English | MEDLINE | ID: covidwho-1911128
ABSTRACT
Companion animals are susceptible to a variety of coronaviruses, and recent studies show that felines are highly susceptible to SARS-CoV-2 infection. RT-PCR diagnostic is currently the method of choice to detect the presence of SARS-CoV-2-specific viral nucleic acids in animal samples during an active infection; however, serological assays are critical to determine whether animals were exposed to the virus and to determine the seroprevalence of SARS-CoV-2-specific antibodies in a defined population. In this study, we utilized recombinant nucleocapsid (N) protein and the receptor-binding domain (RBD) of the spike protein of SARS-CoV-2 expressed in E. coli (N) and mammalian cells (N, RBD) to develop indirect ELISA (iELISA) tests using well-characterized SARS-CoV-2-positive and -negative cat serum panels from previous experimental cat challenge studies. The optimal conditions for the iELISA tests were established based on checkerboard dilutions of antigens and antibodies. The diagnostic sensitivity for the detection of feline antibodies specific for the N or RBD proteins of the iELISA tests was between 93.3 and 97.8%, respectively, and the diagnostic specificity 95.5%. The iELISAs developed here can be used for high-throughput screening of cat sera for both antigens. The presence of SARS-CoV-2-specific antibodies in a BSL-2 biocontainment environment, unlike virus neutralization tests with live virus which have to be performed in BSL-3 laboratories.
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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Observational study Language: English Journal: Front Vet Sci Year: 2022 Document Type: Article Affiliation country: Fvets.2022.864884

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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Observational study Language: English Journal: Front Vet Sci Year: 2022 Document Type: Article Affiliation country: Fvets.2022.864884