Present study revealed application of TaqMan real time PCR technique for diagnosis of rabies viral RNA from 11/20 antemortem skin samples with a sensitivity of 92.85%. The successful use of TaqMan real time PCR concluded that it was a viable, alternative tool for the antemortem diagnosis of rabies. Results correlated well with those of the post-mortem fluorescent antibody test performed on brain specimens of same animals.

No abstract

Aim: Early diagnosis of dreadful rabies is of utmost importance to restrict number of contacts and timely administration of post exposure prophylaxis. The present study was conducted to evaluate the sensitivity comparison of Nested RT-PCR with TaqMan real time PCR technique for intravitam diagnosis of rabies in animals from urine samples. Materials and Methods:Advance molecular approaches Nested RT-PCR and TaqMan real time PCR was employed on 21 urine samples for intravitam diagnosis of rabies. Comparison of both the techniques was done with standard immunofluorescence test (FAT) applied on brain for postmortem confirmation of rabies.Result: Rabies viral RNA was detected in 6/21 (28.57%) and 11/21 (52.38%) urine samples by application of Nested RT-PCR and TaqMan real time PCR respectively. Sensitivity obtained from both the techniques was 62.50% and 78.94% respectively when compared with gold standard immunofluorescence test (FAT). Conclusion:TaqMan real time PCR can serve as more sensitive and viable approach for the intravitam diagnosis of rabies as compared to Nested RT-PCR for detection of rabies from urine of suspected animals. Clinical importance:This study may serve as background for future intravitam rabies diagnostics.

Aim: The present study deals with molecular technique Nested RT-PCR for detection of rabies viral RNA from biological fluid samples (Saliva, Milk and Urine) collected from animal suspected for rabies and to compare the sensitivity of Nested RT-PCR applied for ante mortem diagnosis of rabies with conventional technique (immunofluorescence) applied on neural tissue. Molecular technique Materials and Methods: Nested RT-PCR was applied on 62 biological fluid specimens collected from rabies suspected animals. First round amplification with nested set of primers (RabN1 and RabN5) yielded 1477 bp product while amplification with second round primers (RabNfor and RabNrev) yielded 762 bp product. Sensitivity of the technique was compared in accordance with WHO recommended gold standard test viz. Immunofluorescence (FAT) applied on brain samples. Results: By Nested RT-PCR, viral RNA could be detected in 9/24 (37.50%) saliva samples, 2/17 (11.76%) milk samples and 6/21 (28.57%) urine samples. Confirmatory diagnosis by Immunofluorescence performed on brain sample revealed 18 true positive cases. Overall, Sensitivity of Nested RT-PCR technique employed on fluid samples was 69.23% when compared with immunofluorescence performed on brain samples. Conclusions: Early reliable ante mortem diagnosis of rabies can be obtained from biological fluid samples of animals suspected to be rabid when tested with Nested RT-PCR technique.

: For the productive application of molecular approaches for detection of rabies in live animals 57.89% 57.89%, 78.57% and 52.38% respectively and 3, 3, 6, 1 out of 9 Nested PCR confirmed cases with a sensitivity of 60%, 60%, 52.94% and 75%.