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KMID : 0545120200300030459
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Journal of Microbiology and Biotechnology
2020 Volume.30 No. 3 p.459 ~ p.468
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Rapid and Sensitive Detection of Hepatitis C Virus in Clinical Blood Samples Using Reverse Transcriptase Polymerase Spiral Reaction
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Sun Wenying
Du Ying
Li Xingku
Du Bo
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Abstract
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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.
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KEYWORD
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Hepatitis C virus, polymerase spiral reaction, rapid detection, clinical detection
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