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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.
Index Copernicus ICI Journals Master List 2019 - IJCMAS--ICV 2019: 96.39 For more details click here
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2020)
[Effective from January 1, 2020]
For more details click here

ICV 2019: 96.39
Index Copernicus ICI Journals Master List 2019 - IJCMAS--ICV 2019: 96.39
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 2018: 95.39
NAAS RATING 2020: 5.38

Int.J.Curr.Microbiol.App.Sci.2018.7(8): 1311-1316
DOI: https://doi.org/10.20546/ijcmas.2018.708.148


Purification of Potato Virus Y for Polyclonal Antisera Production and Assessment of Antisera Specificity through DAS-ELISA
Ranima Mishra1*, P.D. Nath1, B. Raigond2, R.C. Boro3, Shankar Hemanta Gogoi1 and Jutimala Phookan1
1Department of Plant Pathology
3Department of Agricultural Biotechnology, Assam Agricultural University, Jorhat 785013, Assam, India
2Division of Plant Protection, ICAR- Central Potato Research Centre (CPRI), Shimla, Himachal Pradesh, India
*Corresponding author
Abstract:

Purified viral protein was obtained from Potato virus Y (PVY) pure culture maintained on potato (Solanum tuberosum L.) plants. The average concentration of the purified materials was found to be 153.1 ng/µl. PVY purified virus preparation was used for immunizing rabbit for production of polyclonal antisera against the virus. Good quality antisera were collected one week post boosters (AS4b, AS5b, AS6b and AS7b) and the IgG fractions were separated through ammonium sulphate precipitation. These four IgG fractions were tested for the detection of PVY by DAS-ELISA with universal anti- rabbit enzyme conjugate as secondary antibody, resulted high specificity with the known PVY infected samples. The assay was compared with the commercial DAS-ELISA kit (Bio Reba, AG, Switzerland). Results revealed that, among the antisera; AS6b was showing the highest mean absorbance values for all positive samples (2.210) and the value shown by the commercial kit was slightly higher i.e., 2.250 and these were followed by AS5b (1.680), AS7b (0.929) and AS4b (0.362), respectively. The absorbance values shown by AS6b were statistically at par with the commercial kit.


Keywords: DAS-ELISA, Purification, Polyclonal antisera, Potato virus Y
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How to cite this article:

Ranima Mishra, P.D. Nath, B. Raigond, R.C. Boro, Shankar Hemanta Gogoi and Jutimala Phookan. 2018. Purification of Potato Virus Y for Polyclonal Antisera Production and Assessment of Antisera Specificity through DAS-ELISA.Int.J.Curr.Microbiol.App.Sci. 7(8): 1311-1316. doi: https://doi.org/10.20546/ijcmas.2018.708.148