Development and evaluation of a novel antigen capture assay for the detection of classical swine fever virus antigens

Veterinary Microbiology
Alfonso ClavijoR A Heckert

Abstract

An antigen-capture enzyme immunoassay (EIA) was developed to detect classical swine fever virus (CSFV) antigen directly from 10% w/v tissue suspension. The assay, based on the sandwich principle, uses a biotinylated monoclonal antibody bound to streptavidin-coated microplates as the capture system and a swine anti-CSFV antibody and rabbit anti-swine HRPO-conjugate as the detector system. The antigen-capture EIA was compared with conventional virus isolation and polymerase chain reaction (PCR) for detection of CSFV in tissues. The ability of the antigen-capture EIA to discriminate classical swine fever (CSF) from bovine viral diarrhea and African swine fever viruses was also tested. The assay was shown to detect 21 different strains of CSFV and was unreactive with tissues from uninfected animals. Signal to noise (S/N) ratios were calculated from the EIA absorbance values. Readings from samples positive by virus isolation (n = 47) averaged a S/N ratio of 5.34. In contrast, samples negative by virus isolation (n = 96) demonstrated a mean S/N ratio of 0.16. At S/N cut-off value of 1.0, all samples that yield virus isolation and PCR negative result were negative in the antigen-capture EIA. Compared with virus propagation in tissue c...Continue Reading

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Citations

Feb 13, 2001·Journal of Microbiological Methods·M R BonenS A Hoffman
Feb 10, 2011·Transboundary and Emerging Diseases·M-L PenrithC Mather
Jan 12, 2005·Clinical and Diagnostic Laboratory Immunology·Min LinJohn Pasick
Jan 22, 2015·Animal Health Research Reviews·Vishal ChanderRishendra Verma
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Dec 4, 2012·Journal of Virological Methods·Bingling YunYulong Gao
Sep 13, 2011·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·August J EberlingTammy R Beckham

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