DEVELOPMENT OF IN-HOUSE ELISA AND ITS COMPARATIVE EVALUATION WITH RFFIT FOR ESTIMATION OF ANTI-RABIES VACCINAL ANTIBODIES IN DOGS
| dc.contributor.advisor | Dr. SHRIKRISHNA ISLOOR | |
| dc.contributor.author | A. K. SANTOSH | |
| dc.date.accessioned | 2021-01-21T06:26:15Z | |
| dc.date.available | 2021-01-21T06:26:15Z | |
| dc.date.issued | 2017-10 | |
| dc.description.abstract | The study was undertaken to develop in-house ELISA and it’s comparision with RFFIT. The recombinant pETRVL-G plasmid having G gene of Dr. Largi’s strain of rabies virus was cloned into pFastBacTM1 vector in DH5α E.coli. The recombinant pFastBacTM1 was confirmed by colony PCR and RE digestion (EcoR I & Sal I) and transposed into DH10 Bac. Recombinant bacmid was selected based on blue-white colonies and confirmed by PCR. Further, the DNA extracted and transformed into Sf-21 cells. The resultant recombinant baculovirus were used to obtain P2 and P3 stock virus and P3 stock used for expression. The expressed recombinant rabies virus G protein was analysed by SDS-PAGE, western blot and doublet bands of 62 and 59 kDa were observed. The crude lysate of baculovirus infected Sf-21 cells was used in ELISA and optimised to arrive at 500ng/100 µl of antigen, 1:100 dilution of serum and 1:15,000 dilution of anti-dog HRPO conjugate. The serum samples from vaccinated dogs (n=247) were tested by RFFIT and in-house ELISA. By RFFIT, 212 serum samples were found to have protective neutralising antibody titre accounting for 85.82 per cent protection. By in-house ELISA , 199 serum samples were found reactive, showing per cent, positivity (PP) more than 36.29 as cut off accounting for 80.56 per cent protection. The diagnostic sensitivity and specificity of in-house ELISA was 90.56 and 80 per cent, respectively with the kappa value of 0.614 revealing good agreement. Usage of purified recombinant rabies virus G protein may be suitable for in-house ELISA in sero-monitoring of antirabies virus neutralising antibodies. | en_US |
| dc.identifier.uri | https://krishikosh.egranth.ac.in/handle/1/5810160489 | |
| dc.keywords | VETERINARY MICROBIOLOGY | en_US |
| dc.language.iso | English | en_US |
| dc.pages | 261 | en_US |
| dc.publisher | KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCIES UNIVERSITY, BIDAR | en_US |
| dc.sub | Veterinary Microbiology | en_US |
| dc.theme | VETERINARY MICROBIOLOGY | en_US |
| dc.these.type | M.V.Sc. | en_US |
| dc.title | DEVELOPMENT OF IN-HOUSE ELISA AND ITS COMPARATIVE EVALUATION WITH RFFIT FOR ESTIMATION OF ANTI-RABIES VACCINAL ANTIBODIES IN DOGS | en_US |
| dc.type | Thesis | en_US |