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경쟁적 역전사-중합효소연쇄반응과 DNA-ELISA법을 이용한 C형 간염 바이러스 RNA 정량
서순팔,김세종,기승정,서강석 대한간학회 2000 Clinical and Molecular Hepatology(대한간학회지) Vol.6 No.2
Background/Aims: 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. Methods: 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. Results: 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). Conclusion: The quantitation of HCV RNA by in-house cRT-PCR and DNA ELISA was more sensitive and had wider range 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)