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Research
(Published
online: 13-01-2015)
11.
Isolation of Canine parvovirus with a
view to identify the prevalent serotype on the basis of partial
sequence analysis -
Gurpreet Kaur, Mudit
Chandra, P. N. Dwivedi and N. S. Sharma
Veterinary World, 8(1): 52-56
doi:
10.14202/vetworld.2015.52-56
Gurpreet Kaur:
Department of Veterinary Microbiology, College of Veterinary
Science, Guru Angad Dev Veterinary and Animal Sciences University,
Ludhiana, Punjab, India; gurpreet7502@rediffmail.com
Mudit
Chandra:
Department of Veterinary Microbiology, College of Veterinary
Science, Guru Angad Dev Veterinary and Animal Sciences University,
Ludhiana, Punjab, India; drmuditchandra@rediffmail.com
P. N.
Dwivedi:
Department of Veterinary Microbiology, College of Veterinary
Science, Guru Angad Dev Veterinary and Animal Sciences University,
Ludhiana, Punjab, India; dwivedi_micro@yahoo.co.in
N. S.
Sharma: Department of Veterinary Microbiology, College of
Veterinary Science, Guru Angad Dev Veterinary and Animal Sciences
University, Ludhiana, Punjab, India;
singhsharman@yahoo.com
Received: 08-09-2014, Revised: 09-12-2014, Accepted: 14-12-2014,
Published online: 13-01-2015
Corresponding author:
Gurpreet Kaur, email: gurpreet7502@rediffmail.com
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.
Keywords: Canine parvovirus, madin darby canine kidney
cell line, polymerase chain reaction, nested polymerase chain
reaction, VP2 gene.
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