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• W2330331207 abstract "Background: In India, control of rabies in dogs by massvaccination is the practical approach. Currently, viable infectious Rabies virus (RABV) dependent and laborious Rapid Fluorescent Focus Inhibition Test (RFFIT) is employed in selected laboratories in India to monitor the vaccinal antibodies. This situation demands an alternative rapid, sensitive, specific and user friendly test. Hence, this study was undertaken to express the recombinant glycoprotein (rRVL-G) of Dr. Larghi's strain of RABV and evaluate it's diagnostic potential. Methods & Materials: The RABV propagated in BHK 21 cells was used as the source of G gene. The Polymerase Chain Reaction (PCR) product of G-gene was cloned into pGEM®-T Easy Vector and transferred into competent Top 10 E. coli. The insert from the recombinants (pGRVL-G) was subcloned into pET32a vector, transferred into TOP 10 cells and recombinants (pETRVL-G) obtained. These recombinants were transferred into E.coli BL21 and screened for expression and immunogenicity by SDS-PAGE and Western Blotting respectively. Indirect ELISA was standardized by Checkerboard titration of complete RABV-G protein (RV-G) and diagnostic potentials of rRVL-G confirmed. Results: The PCR product of RABV complete G gene revealed a band of 1596 bp. The PCR product cloned in pGEM®-T Easy Vector and subbcloned in pET32a vector was confirmed by Restriction Enzyme (RE) digestion and colony PCR (Fig. 1a, 1b) and sequencing. Further SDS-PAGE of the IPTG induced clones revealed a fusionprotein of 75 kDa which was confirmed to be immunogenic by Western blotting (Fig. 2a, 2b) when probed with anti rabies vaccinal dog sera. Standardization of indirect ELISA and application using RV-G and rRVL-G by testing of 40 anti rabies vaccinal dog sera of varying RFFIT titres revealed a significant correlation in the performance of both the antigens (Fig. 3a, 3b).View Large Image Figure ViewerDownload Hi-res image Download (PPT)View Large Image Figure ViewerDownload Hi-res image Download (PPT) Conclusion: The outcome of the work suggested the diagnostic potentials of the recombinant protein in ELISA for seromonitoring of antirabies vaccinal antibodies. In view of the availability of ELISA facilities, expertise and limitations of RFFIT in India, the results encourage the development of recombinant G-protein based ELISA as a newer diagnostics for sero monitoring of antirabies vaccinal antibodies in both domestic and street dogs at a regular interval of time." @default.
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• W2330331207 date "2016-04-01" @default.
• W2330331207 modified "2023-09-24" @default.
• W2330331207 title "Expression of glycoprotein gene of Rabies virus and evaluation of recombinant protein for seromonitoring of vaccinal antibodies in dogs" @default.
• W2330331207 doi "https://doi.org/10.1016/j.ijid.2016.02.075" @default.
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