Rapid and sensitive detection of hepatitis C virus in clinical blood samples using reverse transcriptase polymerase spiral reaction.

Abstract

The aim of this study was to establish a new polymerase spiral reaction (PSR) combining with reverse transcription reactions for HCV detection targeting 5 UTR gene. To avoid cross-contamination of aerosols, an isothermal amplification tube (IATs) as a separate containment control is used for the judgment of the result. After optimizing the RT-PSR reaction system, the effectiveness and specificity of RT-PSR were tested against 15 different virus strains which include 8 HCV positive and 7 non-HCV controls. The results showed that RT-PSR assay effectively detected all 8-HCV strains, and no false positive were found in 7 non-HCV strains. The detection limit of RT-PSR assay was comparable to the real-time RT-PCR, but was more sensitive than the RT-LAMP. The established RT-PSR assay was further evaluated for detection of HCV in clinical blood samples, the results of 80.25% of detection rate by RT-PSR demonstrated a better or similar effectiveness compared to the RT-LAMP (79.63%) and real-time RT-PCR (80.25%). The results demonstrated that the RT-PSR assay offers high specificity, sensitivity for HCV detection and has great potential for screening HCV in clinical blood samples.