Isolation of Canine parvovirus with a view to identify the prevalent serotype on the basis of partial sequence analysis.

Gurpreet Kaur,P N Dwivedi,Mudit Chandra,N S Sharma

doi:10.14202/vetworld.2015.52-56

Abstract

Aim:The aim of this study was to isolate Canine parvovirus (CPV) from suspected dogs on madin darby canine kidney (MDCK) cell line and its confirmation by polymerase chain reaction (PCR) and nested PCR (NPCR). Further, VP2 gene of the CPV isolates was amplified and sequenced to determine prevailing antigenic type.Materials and Methods:A total of 60 rectal swabs were collected from dogs showing signs of gastroenteritis, processed and subjected to isolation in MDCK cell line. The samples showing cytopathic effects (CPE) were confirmed by PCR and NPCR. These samples were subjected to PCR for amplification of VP2 gene of CPV, sequenced and analyzed to study the prevailing antigenic types of CPV.Results:Out of the 60 samples subjected to isolation in MDCK cell line five samples showed CPE in the form of rounding of cells, clumping of cells and finally detachment of the cells. When these samples and the two commercially available vaccines were subjected to PCR for amplification of VP2 gene, a 1710 bp product was amplified. The sequence analysis revealed that the vaccines belonged to the CPV-2 type and the samples were of CPV-2b type.Conclusion:It can be concluded from the present study that out of a total of 60 samples 5 samples exhibited CPE as observed in MDCK cell line. Sequence analysis of the VP2 gene among the samples and vaccine strains revealed that samples belonged to CPV-2b type and vaccines belonging to CPV-2.

Highlights

Diarrhea is a very common condition in dogs caused by various viral and bacterial causative agents
The sequence analysis revealed that the vaccines belonged to the Canine parvovirus (CPV)-2 type and the samples were of CPV-2b type
It can be concluded from the present study that out of a total of 60 samples 5 samples exhibited cytopathic effects (CPE) as observed in madin darby canine kidney (MDCK) cell line

Summary

Introduction

Diarrhea is a very common condition in dogs caused by various viral and bacterial causative agents. It has economic implications when a large number of dogs are affected in quick succession [1]. It is multifactorial and the origin of Canine parvovirus (CPV) though is not absolutely clear but phylogenetically originated from feline panleukopenia virus or a very closely related carnivore parvovirus of feral canids like foxes and mink [2]. CPV is prone to genetic evolution and has undergone several mutations that have led to various antigenic variants of CPV-2 that have replaced the original CPV-2 [4]. CPV-2 can infect both canine and feline cells in vitro but infects only dogs in vivo [2]

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