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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
IJCMAS is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCMAS Articles.
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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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Original Research Articles

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.2018.7(4): 2837-2844
DOI: https://doi.org/10.20546/ijcmas.2018.704.323


Plaque Purification of Bluetongue Virus Serotype-4 (BTV-4)
M. Srikanth Reddy1, Kalyani Putty1*, Y. Narasimha Reddy1, P.P. Rao2, M.Ramakoti Reddy3, Uma2, Sunil R. Patil1, Susmitha Birru1, Abdul Muzeer Shaik1 and Dhanalakshmi Kancharlapally1
1College of Veterinary Science, Rajendranagar, Hyderabad-500030, Telangana, India
2Ella Foundation, Genome Valley Hyderabad-500 078, Telangana, India
3Directorate of Poultry Research, Rajendranagar, Hyderabad-500030, Telangana, India
*Corresponding author
Abstract:

The present study was taken up with a view to undertake plaque purification of BTV-4 isolate available in the department, to confirm its purity. BTV-4 isolate was adapted to Vero cell line and had a titer of 106.46/ml after 8 passages in the process of plaque purification which was determined as 104.833/ml prior to adaption. Plaque purification was carried for three times using agarose overlaying method in 6 well plate. Each time plaques were observed from 4th day of infection of approximately not more than 1mm diameter. One of the plaques from third round of purification was grown in T25 flask and the resultant cell culture fluid was used for RNA isolation and RT-PCR through which the respective plaque was confirmed as BTV-4. Therefore, it was confirmed that there was no other serotype other than BTV-4 and the plaque purified virus can be used either to study pathogenesis or to raise hyperimmune serum.


Keywords: BTV-4, Vero cells, TCID50, Plaque purification, Sea plaque agarose, RT-PCR
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How to cite this article:

Srikanth Reddy M., Kalyani Putty, Y. Narasimha Reddy, P.P. Rao, M. Ramakoti Reddy, Uma, Sunil R. Patil, Susmitha Birru, Abdul Muzeer Shaik and Dhanalakshmi Kancharlapally. 2018. Plaque Purification of Bluetongue Virus Serotype-4 (BTV-4).Int.J.Curr.Microbiol.App.Sci. 7(4): 2837-2844. doi: https://doi.org/10.20546/ijcmas.2018.704.323