Rapid and Sensitive Detection of Hepatitis C Virus in Clinical Blood Samples Using Reverse Transcriptase Polymerase Spiral Reaction

• Published in: Journal of microbiology and biotechnology Vol. 30; no. 3; pp. 459 - 468
• Main Authors: Sun, Wenying, Du, Ying, Li, Xingku, Du, Bo
• Format: Journal Article
• Language: English
• Published: Korea (South) Korean Society for Microbiology and Biotechnology 28.03.2020; 한국미생물·생명공학회
• Subjects: Hepacivirus - genetics; Hepacivirus - isolation & purification; Hepatitis C - virology; Humans; Research article; Reverse Transcriptase Polymerase Chain Reaction; RNA, Viral - genetics; Time Factors; 생물학; clinical detection; rapid detection; Hepatitis C virus; polymerase spiral reaction
• ISSN: 1017-7825; 1738-8872
• DOI: 10.4014/jmb.1910.10041

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