실시간 정량적 역전사-중합효소연쇄반응을 이용한 HCV 정량검사 시약(Real-Q HCV Quantification Kit)의 평가
Evaluation of the Real-Q HCV Quantification Kit
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배경: 정확한 Hepatitis C Virus (HCV) RNA의 정량은 치료반응 예측과 치료결과 평가에 매우 중요한 지표로 활용된다. 최근 HCV RNA 정량에 사용되고 있는 실시간 정량적 역전사-중합효소연쇄반응(real-time PT-PCR)은 측정 범위가 넓고 결과값의 직선성이 좋아 기존에 사용되던 branched DNA법과 PCR-hybridization법을 대체하고 있다. 본 연구에서는 국내 에서 개발된 Real-Q HCV Quantification kit의 성능을 평가하였다. 방법: Real-Q HCV Quantification kit의 성능평가를 위해 kit의 검출한계, 분석특이도, 정밀도, 직선성 범위, HCV RNA 표준 물질의 회수율을 평가하였고, Cobas Amplicor HCV Monitor 2.0과의 상관성을 분석하였다. 결과: 다양한 농도의 HCV RNA 표준물질에 대하여 높은 회수율을 나타내었고, 모든 유전자형에서 동일하게 증폭되는 것을 확인하였다. Hepatitis B virus와 Human Immunodeficiency virus 양성 검체와 교차반응이 없음을 확인하였고, 검사 중 변이계수는 9.52∼15.84%, 검사간 변이계수는 9.40∼17.53% 범위이고 일간변이계수(between-day coefficient of variation) 는 11.62∼18.04%를 보였으며, 검출한계는 44 IU/mL이었다. Cobas Amplicor HCV Monitor 2.0과도 좋은 상관성을 보 였다. 결론: 본 연구를 통하여 평가된 Real-Q HCV Quantification kit는 특이도, 민감도, 직선성, 정밀도에서 우수한 성능을 보여, HCV RNA 정량을 이용한 만성 C형 간염 치료의 추적관찰에 유용하게 사용될 수 있을 것으로 판단된다.
Abstract
Background: Hepatitis C virus (HCV) RNA quantification is necessary for predicting the therapeutic response and assessing treatment results in patients with chronic HCV infection. Recently, real-time PCR technology for HCV RNA quantification displayed good linearity within the dynamic range. Thus, it is gradually replacing branched-DNA (bDNA) and PCRhybridization assays. In this study, we evaluated the performance of the Real-QTM HCV quantification kit (biosewoom. Inc., Seoul, Korea) developed in Korea. Methods: We evaluated the HCV quantification kit for detection limit, specificity, linearity, accuracy, and recovery rate of HCV RNA standard material. The results were analyzed for a correlation with those of Cobas Amplicor HCV Monitor 2.0. Results: The HCV quantification kit showed a high recovery rate of HCV RNA standard material of various concentrations and amplication of HCV RNA equally in all genotypes. Hepatitis B virus and human immunodeficiency virus showed no cross-reactivity with HCV. Within-run and between-run coefficients of variation (CV) were 9.52∼15.84% and 9.40∼17.53%, respectively. Between-day coefficients of variation were 11.62∼18.04%, and detection limit was 44 IU/mL. It showed a good correlation with Cobas Amplicor HCV Monitor 2.0 (R2=0.8954). Conclusion: The Real-Q HCV quantification kit showed a good specificity, sensitivity, linearity, and accuracy; therefore, we propose that it is fully adequate for monitoring antiviral therapy in patients with chronic HCV infection.
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