Abstract
Aim:The present study was conducted to detect the presence of canine parvovirus (CPV) among diarrheic dogs in Himachal Pradesh and to identify the most prevalent antigenic variant of CPV based on molecular typing and sequence analysis of VP2 gene.Materials and Methods:A total of 102 fecal samples were collected from clinical cases of diarrhea or hemorrhagic gastroenteritis from CPV vaccinated or non-vaccinated dogs. Samples were tested using CPV-specific polymerase chain reaction (PCR) targeting VP2 gene, multiplex PCR for detection of CPV-2a and CPV-2b antigenic variants, and a PCR for the detection of CPV-2c. CPV-2b isolate was cultured on Madin-Darby canine kidney (MDCK) cell lines and sequenced using VP2 structural protein gene. Multiple alignment and phylogenetic analysis was done using ClustalW and MEGA6 and inferred using the Neighbor-Joining method.Results:No sample was found positive for the original CPV strain usually present in the vaccine. However, about 50% (52 out of 102) of the samples were found to be positive with CPV-2ab PCR assay that detects newer variants of CPV circulating in the field. In addition, multiplex PCR assay that identifies both CPV-2ab and CPV-2b revealed that CPV-2b was the major antigenic variant present in the affected dogs. A PCR positive isolate of CPV-2b was adapted to grow in MDCK cells and produced characteristic cytopathic effect after 5th passage. Multiple sequence alignment of VP2 structural gene of CPV-2b isolate (Accession number HG004610) used in the study was found to be similar to other sequenced isolates in NCBI sequence database and showed 98-99% homology.Conclusion:This study reports the first detection of CPV-2b in dogs with hemorrhagic gastroenteritis in Himachal Pradesh and absence of other antigenic types of CPV. Further, CPV-specific PCR assay can be used for rapid confirmation of circulating virus strains under field conditions.
Highlights
Canine parvoviral infection is caused by canine parvovirus 2 (CPV-2) of family Parvoviridae
About 50% (52 out of 102) of the samples were found to be positive with CPV-2ab polymerase chain reaction (PCR) assay that detects newer variants of CPV circulating in the field
A PCR positive isolate of CPV-2b was adapted to grow in Madin-Darby canine kidney (MDCK) cells and produced characteristic cytopathic effect after 5th passage
Summary
Canine parvoviral infection is caused by canine parvovirus 2 (CPV-2) of family Parvoviridae. CPV-2 is a non-enveloped single-stranded DNA virus, 5.2 kb in size, has icosahedral symmetry and encodes for two structural (capsid) proteins (VP1 and VP2) and two non-structural proteins (NS1 and NS2). The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated
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