Development of a polyclonal antibody-based AC-ELISA and its comparison with PCR for diagnosis of canine parvovirus infection.
A polyclonal antibody-based antigen-capture ELISA (AC-ELISA) has been developed for detection of Canine parvovirus (CPV) antigens in faecal samples of dogs. The assay uses rabbit anti-CPV polyclonal antibody as the capture antibody, guinea pig anti-CPV polyclonal antibody as tracing antibody and anti-guinea pig HRPO conjugate as the detection system. The optimum dilution of the capture antibody and the tracing antibody capable of detecting the CPV-2 antigens was found to be 1:1 600 and 1:400, respectively, in the check-board titration. In this study, a total of 152 samples (129 faecal samples and 23 cell culture supernatant) were tested both by AC-ELISA and by polymerase chain reaction (PCR). Of the samples tested, 69 and 78 samples were found positive by AC-ELISA and PCR, respectively. The AC-ELISA had relative sensitivity, relative specificity and accuracy of 88.4%, 100.0% and 91.4% respectively. The analytical sensitivity of AC-ELISA was estimated to be 10(2.8) TCID(50)/mL whereas PCR sensitivity was 10(0.8) TCID(50)/mL. The AC-ELISA is a simple, quick and reliable method for screening large numbers of faecal samples of dogs suspected of CPV infection.
Kumar
Manoj
M
Virology Laboratory, Centre for Animal Disease Research and Diagnosis, Indian Veterinary Research Institute, Izatnagar, Bareilly, U.P., 243122, India.
Nandi
Sukdeb
S
Chidri
Sunil
S
D000818:Animals; D000914:Antibodies, Viral; D004283:Dog Diseases / Q000175:diagnosis* / Q000821:virology; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005243:Feces / Q000821:virology; D006168:Guinea Pigs; D010322:Parvoviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D017993:Parvovirus, Canine / Q000276:immunology / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
D000914:Antibodies, Viral