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 DH5 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 ul 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.