Canine parvovirus 2 (CPV-2) is the causative agent of acute hemorrhagic enteritis and myocarditis in dogs. In this study the nucleotide sequence of the VP1/VP2 gene of a CPV isolate from India was analyzed and the phylogenetic relationship with other CPV isolates was established. Out of 36 samples analyzed, 16 were found positive for CPV-2 by polymerase chain reaction (PCR). Among the 16 positive samples, five were inoculated in MDCK cells for isolation out of which one adapted successfully to the cell culture system. Phylogenetic analysis based on the nucleotide sequence of the VP-1/VP-2 gene revealed that the Indian isolate closely resembled a CPV-2b Italian strain, showing 98.4% nucleotide sequence homology indicating very little genetic divergence since it was first identified in 1978.
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.
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