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

• Published in: Journal of microbiology and biotechnology Vol. 15; no. 3; pp. 595 - 602
• Main Authors: Kim, B.G. (Korea Food and Drug Administration, Seoul, Republic of Korea), Jeong, H.S. (Korea Food and Drug Administration, Seoul, Republic of Korea), Baek, S.Y. (Korea Food and Drug Administration, Seoul, Republic of Korea), Shin, J.H. (Korea Food and Drug Administration, Seoul, Republic of Korea), Kim, J.O. (Korea Food and Drug Administration, Seoul, Republic of Korea), Min, K.I. (Korea Food and Drug Administration, Seoul, Republic of Korea), Ryu, S.R. (Korea Food and Drug Administration, Seoul, Republic of Korea), Min, B.S. (Korea Food and Drug Administration, Seoul, Republic of Korea), Kim, D.K. (Korea Food and Drug Administration, Seoul, Republic of Korea), Jeong, Y.S. (Kyung Hee University, Seoul, Republic of Korea), Park, S.N. (Korea Food and Drug Administration, Seoul, Republic of Korea), E-mail: suenie@kfda.go.kr
• Format: Journal Article
• Language: English
• Published: Seoul Korean Society for Applied Microbiology 01.06.2005
• Subjects: ARN; Automated RNA extraction; Biological and medical sciences; Biotechnology; Fundamental and applied biological sciences. Psychology; HCV RNA; quantitation; real-time RT-PCR; RNA; Automation; RNA; Real time; real-time RT-PCR; Virus; Polymerase chain reaction; Rapid technique; Extraction; Flaviviridae; Detection; Hepatitis C virus; Hepacivirus; Quantitative analysis; Automated RNA extraction; HCV RNA. quantitation
• ISSN: 1017-7825

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∨7-10¹ HCV RNA copies/reaction (R²=0.997). The synthetic HCV RNA standard of 1.84±0.1 (mean±SD) copies developed in this study corresponded to 1 international unit (IU) of WHO International Standard tor HCV RNA (96/790 I).