Abstract
From the reasons that canine coronavirus (CCV) grows more efficiently than feline coronavirus in a cell culture and they are mutually related in their antigenicities, an enzyme-linked immunosorbent assay (ELISA) using CCV-infected feline kidney (CRFK) cells as substrate antigens was developed for detection of anti-coronavirus antibodies in cats. It was indispensable for generating coronavirus-specific ELISA antibody activities that the sample was applied to the mock-infected, normal CRFK cells in parallel with the CCV-infected cells and then the optical density values given by the mock-infected cell antigen were subtracted from those given by the virus-infected cell antigen. On the basis of ELISA antibody titers obtained in sera from the cats experimentally infected with CCV and from the spontaneous feline infectious peritonitis (FIP) cases, the ELISA described in the present study was found to be applicable as a simple and easy serologic test which was able to detect anti-coronavirus antibodies as efficiently as the indirect immunofluorescence assay with homologous FIP virus.
Keywords: Coronavirus, feline infectious peritonitis virus, canine coronavirus, enzyme-linked immunosorbent assay, ELISA
Abbreviations: CCV, canine coronavirus; CRFK, Crandell feline kidney; DK/P, primary dog kidney; Eagle's MEM, Eagle's minimal essential medium; ELISA, enzyme-linked immunosorbent assay; FECV, feline enteric coronavirus; FIP, feline infectious peritonitis; FIPV, FIP virus; IFA, indirect immunofluorescence assay; mAb, monoclonal antibody; MNT, micro-neutralization test; NUS, NU-serum; OD, optical density; PFU, plaque forming unit; PBS, phosphate buffered saline solution; TGEV, transmissible gastroenteritis virus; VNT, virus neutralization test
Résumé
Etant donné que coronavirus canin (CVC) prolifère en culture cellulaire avec une meilleure efficacité que coronavirus félin et qu'ils ont une antigénéciteoroisée, un test ELISA utilisant des cellules rénales félines (CRFK) infectées par le CVC comme antigène substrat a été dévélopée pour la détéction des anticorps anti-coronaviraux chez le chat. Pour obtenir une activité anticorps ELISA coronavirus-spécifique, il a été considéré comme indispensable que l'échantillon soit appliqué aux cellules (CRFK) normales, non inféctées en parallèle avec les cellules mentionnées ci-dessus et que les valeurs de densité optique données par l'antigéne des cellules noninfectées soient soustraites à celles donnéees par l'antigène des cellules infectées. En se basant sur les titres d'anticorps ELISA obtenus avec les sérums de chats infectés expérimentalement avec le CVC ou atteiuts de péritonite infectieuse féline (PIF) spontanée, l'ELISA décrit dans cette étude s'est révélé applicable comme un test sérologique à la fois simple et maniable qui nous permet de détécter anticorps anti-coronavirus aussi effectivement que l'essai d'immunofluorescence indirecte avec le virus PIF homoloque.
Mots-clé: Coronavirus, virus de la péritonite infectieuse féline, coronavirus canin, test immunopenzymatique, ELISA
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