The Study of Amplifing HCV ssRNA by RT-PCR Combined With Transcription in vitro

The purpose of this study is to establish a new method which HCV RNA can be used to detect sensitively and specifically through efficient single strand RNAs(ssRNAs) amplification. The sera from the chronic Hepatitis C patients were used as specimens. Single step reverse transcription-PCR (SRT-PCR) was performed prospectively, with a designed set of primers which defined the sequence unique to the 5’ noncoding region of the HCV genome, a great deal of specific ssRNAs were synthesized in the presence of T7RNA polymerase and other appropriate conditions within five hours. Comparing with the nested-PCR, the quantity of the products in our method was nearly 20-fold higher and its specificity was verified by the electrophoresis on agarose gel and the dot hybridization with the probe.