Preliminary report on a single-tube, non-interrupted reverse transcription-polymerase chain reaction for the detection of rabies virus in brain tissue

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.