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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2017: 100.00
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2017: 100.00
For more details click here

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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com / submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2017: 100.00
NAAS RATING 2018: 5.38

Int.J.Curr.Microbiol.App.Sci.2016.5(5): 757-766
DOI: http://dx.doi.org/10.20546/ijcmas.2016.505.077


Detection of Local Isolates of Canine Distemper Virus by Reverse-Transcription Polymerase Chain Reaction
Swati1*, Dipak Deka2 and Ramneek2
1CSIR-Institute of Genomics & Integrative Biology, New Delhi-110020, India
2School of Animal Biotechnology, GADVASU, Ludhiana-141004, India
*Corresponding author
Abstract:

Distemper is a disease of several canids caused by canine distemper virus (CDV).  The present study aimed to detect CDV directly from the swab samples of the suspected dogs by diagnostic RT-PCR. The positive samples were then subjected to virus isolation in MDCK cells after growing it in mitogen stimulated dog lymphocytes and the presence of the viral RNA again was confirmed by RT-PCR. Further, haemagglutinin (H) and largeprotein (L) genes were amplified and cloned into pGEMT Easy cloning vector, sequenced and phylogenetically characterized with the available reference CDV strains in GeneBank. Three samples were detected CDV positive by RT-PCR from a total of 50 samples and virus could be successfully isolated from one sample in MDCK cells after lymphocyte culture. After a number of serial passages, there were no consistent CPE observed but viral RNA could be detected from all the cell culture harvests of all passages which confirmed the presence of CDV. Mitogen stimulated dog lymphocyte culture proved to be an effective and economical method to isolate wild-type CDV from clinical samples. Partial haemagglutinin (H) and large protein (L) gene sequences of one of the CDV isolates made a distinct clade in phylogenetic tree which was clearly separated from commercial CDV vaccine groups (Lederle, Ondersteport, Convac and Synder Hill).


Keywords: Canine distemper virus, RT-PCR, isolation, viral passages, molecular characterization
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How to cite this article:

Swati, Dipak Deka and Ramneek. 2016. Detection of Local Isolates of Canine Distemper Virus by Reverse-Transcription Polymerase Chain ReactionInt.J.Curr.Microbiol.App.Sci. 5(5): 757-766. doi: http://dx.doi.org/10.20546/ijcmas.2016.505.077