Pk. Gupta et al., Preliminary report on a single-tube, non-interrupted reverse transcription-polymerase chain reaction for the detection of rabies virus in brain tissue, VET RES COM, 25(3), 2001, pp. 239-247
A simple method for the rapid detection of rabies virus was developed emplo
ying a single-tube reverse transcription-polymerase chain reaction (RT-PCR)
. The method utilized a single buffer system for both RT and PCR and was pe
rformed without interruption as a single thermal cycling programme. Two pri
mer sets within the genes coding for rabies nucleoprotein and glycoprotein
were used to amplify a 533 bp and a 406 bp amplicon, respectively. The ampl
ified products were detected with a challenge virus strain (CVS) of rabies.
There was no amplified product with uninfected mouse or dog brain. The met
hod was applied to detect rabies virus in 10 mouse inoculation test (MIT)-p
ositive and three MIT-negative brain tissue samples. The amplified product
was found only in the MIT-positive samples. The amplified product was confi
rmed by restriction endonuclease analysis using Hinf1. The results from RT-
PCR correlated well with the results from MIT. This indicates that the sing
le-tube RT-PCR may be a useful method for detecting rabies virus in brain t
issue samples from suspected cases of rabies.