title: First detection of canine parvovirus type 2b from diarrheic dogs in Himachal Pradesh.
abstract: OBJECTIVE
  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.
  
  
  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 5thpassage. 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Veterinary Microbiology, Dr. G. C. Negi College of Veterinary and Animal Sciences, Chaudhary Sarwan Kumar Himachal Pradesh Krishi Vishvavidyalaya, Palampur - 176 062, Himachal Pradesh, India.
authors.forename: Shalini
authors.initials: S
authors.lastname: Sharma
authors.affiliation: Department of Veterinary Microbiology, Dr. G. C. Negi College of Veterinary and Animal Sciences, Chaudhary Sarwan Kumar Himachal Pradesh Krishi Vishvavidyalaya, Palampur - 176 062, Himachal Pradesh, India.
authors.forename: Prasenjit
authors.initials: P
authors.lastname: Dhar
authors.affiliation: Department of Veterinary Microbiology, Dr. G. C. Negi College of Veterinary and Animal Sciences, Chaudhary Sarwan Kumar Himachal Pradesh Krishi Vishvavidyalaya, Palampur - 176 062, Himachal Pradesh, India.
authors.forename: Aneesh
authors.initials: A
authors.lastname: Thakur
authors.affiliation: Department of Microbiology, College of Basic Sciences, Chaudhary Sarwan Kumar Himachal Pradesh Krishi Vishvavidyalaya, Palampur - 176 062, Himachal Pradesh, India.
authors.forename: Vivek
authors.initials: V
authors.lastname: Sharma
authors.affiliation: Department of Veterinary Microbiology, Dr. G. C. Negi College of Veterinary and Animal Sciences, Chaudhary Sarwan Kumar Himachal Pradesh Krishi Vishvavidyalaya, Palampur - 176 062, Himachal Pradesh, India.
authors.forename: Mandeep
authors.initials: M
authors.lastname: Sharma
keywords: canine parvovirus; molecular typing; phylogenetic analysis; sequencing