TAILIEUCHUNG - Purification of potato virus y for polyclonal antisera production and assessment of antisera specificity through DAS-ELISA

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 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. | Purification of potato virus y for polyclonal antisera production and assessment of antisera specificity through DAS-ELISA 2018 7 8 1311-1316 International Journal of Current Microbiology and Applied Sciences ISSN 2319-7706 Volume 7 Number 08 2018 Journal homepage http Original Research Article https Purification of Potato Virus Y for Polyclonal Antisera Production and Assessment of Antisera Specificity through DAS-ELISA Ranima Mishra1 . Nath1 B. Raigond2 . Boro3 Shankar Hemanta Gogoi1 and Jutimala Phookan1 1 Department of Plant Pathology 3Department of Agricultural Biotechnology Assam Agricultural University Jorhat 785013 Assam India 2 Division 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 Keywords potato Solanum tuberosum L. plants. The average concentration of the purified materials was found to be ng µl. PVY purified virus preparation was used for immunizing DAS-ELISA rabbit for production of polyclonal antisera against the virus. Good quality antisera were Purification collected one week post boosters AS4b AS5b AS6b and AS7b and the IgG fractions Polyclonal antisera were separated through ammonium sulphate precipitation. These four IgG fractions were Potato virus Y tested for the detection of PVY by DAS-ELISA with universal anti- rabbit enzyme Article Info 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 Accepted Switzerland . Results revealed that among the antisera AS6b was showing the highest 08 July 2018 mean absorbance values for all positive samples and the value shown by the Available Online commercial kit was slightly higher . and these were followed by AS5b 10 .

• Nông nghiệp
• DAS ELISA
• Polyclonal antisera
• Potato virus Y
• Antisera specificity through DAS ELISA
• Polyclonal antisera production