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Korean J Hepatol > Volume 6(2); 2000 > Article
The Korean Journal of Hepatology 2000;6(2): 156-171.
원저 : 경쟁적 역전사-중합효소연쇄반응과 DNA-ELISA법을 이용한 C형 간염 바이러스 RNA 정량 ( Quantitation of Hepatitis C Virus RNA by Competitive RT-PCR and DNA-ELISA Method )
Quantitation of Hepatitis C Virus RNA by Competitive RT-PCR and DNA-ELISA Method
Kang Seok Seo, M.D., Seung Jung Kee1, M.D., Soon Pal Suh1, M.D., Sei Jong Kim, M.D.
Department of Internal Medicine and Clinical Pathology, Chonnam University Medical School, Kwangju, Korea

Quantitation of Hepatitis C Virus (HCV) RNA in serum is important for monitoring the response to interferon-α therapy in patients with chronic hepatitis C. Several commercial assays are recently available, but they are expensive and cannot be used as a gold standard.
An in-house competitive reverse transcription-polymerase chain reaction (cRT-PCR) was developed and validated. The procedure involves the construction of a mutant and wild type HCV RNA internal standard (IS), cRT-PCR, and colorimetric detection with DNA-ELISA. A standard curve was obtained and used for final HCV RNA quantitation.
The standard curve was linear over the range of 1×104 to 5×107 copies/mL of the HCV RNA standard (r=0.976). This in-house cRT-PCR was comparable with the branched DNA (bDNA) assay (Quantiplex HCV 2.0, Chiron, USA) with positive correlation between the two tests (r=0.735).
The quantitation of HCV RNA by in-house cRT-PCR and DNA ELISA was more sensitive and had wider range of detection compared to bDNA assay. This assay is useful for follow-up of HCV RNA concentration after interferon-α therapy.(Korean J Hepatol 2000;6:156-171)
KeyWords: Hepatitis/Viral/Hepatitis C, Quantitation of Hepatitis C Virus RNA
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