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Journal of microbiology and biotechnology v.15 no.3, 2005년, pp.595 - 602   피인용횟수: 1

Validation of One-Step Real-Time RT-PCR Assay in Combination with Automated RNA Extraction for Rapid Detection and Quantitation of Hepatitis C Virus RNA for Routine Testing in Clinical Specimens

KIM BYOUNG-GUK    (Department of Biologics Evaluation, Korea Food and Drug Administration   ); JEONG HYE-SUNG    (Department of Biologics Evaluation, Korea Food and Drug Administration   ); BAEK SUN-YOUNG    (Department of Biologics Evaluation, Korea Food and Drug Administration   ); SHIN JIN-HO    (Department of Biologics Evaluation, Korea Food and Drug Administration   ); KIM JAE-OK    (Department of Biologics Evaluation, Korea Food and Drug Administration   ); MIN KYUNG-IL    (Department of Biologics Evaluation, Korea Food and Drug Administration   ); RYU SEUNG-REL    (Department of Biologics Evaluation, Korea Food and Drug Administration   ); MIN BOK-SOON    (Department of Biologics Evaluation, Korea Food and Drug Administration   ); KIM DO-KEUN    (Department of Biologics Evaluation, Korea Food and Drug Administration   ); JEONG YONG-SEOK    (Molecular Virology Laboratory, Department of Biology, Kyung-Hee University   ); PARK SUE-NIE    (Department of Biologics Evaluation, Korea Food and Drug Administration  );
  • 초록

    A one-step real-time quantitative RT-PCR assay in combination with automated RNA extraction was evaluated for routine testing of HCV RNA in the laboratory. Specific primers and probes were developed to detect 302 bp on 5'-UTR of HCV RNA. The assay was able to quantitate a dynamic linear range of $10^7-10^1$ HCV RNA copies/reaction ( $R^2=0.997$ ). The synthetic HCV RNA standard of $1.84{\pm}0.1\;(mean{\pm}SD)$ copies developed in this study corresponded to 1 international unit (IU) of WHO International Standard for HCV RNA (96/790 I). The detection limit of the assay was 3 RNA copies/reaction (81 IU/ml) in plasma samples. The assay was comparable to the Amplicor HCV Monitor (Monitor) assay with correlation coefficient r=0.985, but was more sensitive than the Monitor assay. The assay could be completed within 3 h from RNA extraction to detection and data analysis for up to 32 samples. It allowed rapid RNA extraction, detection, and quantitation of HCV RNA in plasma samples. The method provided sufficient sensitivity and reproducibility and proved to be fast and labor-saving, so that it was suitable for high throughput HCV RNA test.


  • 주제어

    Automated RNA extraction .   HCV RNA .   quantitation .   real-time RT-PCR.  

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  • 이 논문을 인용한 문헌 (1)

    1. 2008. "" Journal of microbiology and biotechnology, 18(11): 1858~1861     

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