SEPARATE DETECTION OF THE 2 COMPLEMENTARY RNA STRANDS OF HEPATITIS-A VIRUS

The minus strand of hepatitis A virus can be detected specifically by reverse transcription and polymerase chain reaction amplification in i nfected cell culture extracts. Several controls gave evidence that the amplified fragment actually used the minus strand as initial template . Non-thermostable reverse transcriptase was not efficient for this pu rpose because of self-priming of the positive-stranded viral RNA durin g the reverse transcription step. This problem was overcome by the use of the thermostable rTth DNA polymerase that also has reverse transcr iptase activity in the presence of Mn2+.