실시간 정량적 중합효소연쇄반응을 이용한 Real-Q HBV Quantification Kit의성능평가 (Performance Evaluation of Real-Q HBV Quantification Kit for HBV DNA by Real-Time PCR)

서지정보

· 발행기관 : 대한진단검사의학회
· 수록지 정보 : Annals of Laboratory Medicine / 26권 / 6호 / 442 ~ 448페이지
· 저자명 : 황상현, 차충환, 김유리, 권오중, 오흥범

초록

배경 : Hepatitis B virus (HBV) DNA 정량검사는 항바이러스제제의 사용, 치료효과 추적, 내성 HBV의 조기 진단 등 HBV감염의 치료를 위해 필요하다. 최근 검사실에 보급되고 있는 real-time quantitative PCR은 기존의 HBV DNA 검사법의 단점들을 보완하면서 보다 간편하고 정확한 HBV DNA 정량 기법이다.본 연구에서는 최근 국내에서 개발된 Real-Q HBV Quantifica-tion kit의 성능을 평가하였다.방법 : Real-Q HBV Quantification kit의 회수율, 검출 한계,분석특이도 및 진단특이도, 정밀도, 직선성 범위, 그리고 DigeneHybrid-Capture II와 상관성과 internal control (IC)의 간섭영향을 분석하였다.결과 : 평균 %회수율은 108.6% 이었으며, 동일 검체에서 IC과의 경쟁 때문에 target 유전자의 증폭이 억제되지 않음을 확인하였다. Hepatitis C virus (HCV), human immunodeficiency virus(HIV), Cytomegalovirus (CMV) 19개의 양성 검체와 교차반응은 없었으며, 본 검사의 재현성은 검사차례내 변이계수 8.7-11.9%,검사차례간 변이계수 10.5-14.7%, 검사일간 변이계수 13.2-21.4%를 보였다. 검출 한계는 56 copies/mL이었으며, 보고범위는 102-1× 1010 copies/mL이었다, HBV 음성 100검체를 이용하여 분석한 진단특이도는 100%이었고, Digene Hybrid-Capture II와 우수한 상관관계를 보였다(R2=0.9827).

영어초록

Background : Hepatitis B virus (HBV) DNA quantification is important for the management of HBV infection and identification of the development of resistance. The susceptibility to contamination and more variable reproducibility of results with the conventional HBV DNA quantification method have raised the need of a more simple and accurate method for HBV DNA quantification. Real-time quantitative PCR assays recently introduced in the laboratory can meet these needs. In this study, we evaluated the performance of the Real-Q HBV Quantification kit developed in Korea. Methods : We evaluated the recovery of DNA extraction, the interference of internal control, an analytical sensitivity, specificity, and reproducibility, a clinical specificity, and a reportable range of the Real-Q HBV Quantification kit. The quantification result was also compared to that obtained by the Digene Hybrid-Capture II. Results : The mean percent recovery was 108.6% and there was no interference with the internal control on DNA extraction. None of HIV, hepatitis C virus, or cytomegalovirus showed a cross-reactivity with HBV. This assay detected HBV DNA in a linear range from 102 to 1010 copies/mL, with the detection limit of 56 copies/mL. The assay exhibited a low within-run CV (coefficient of variation) (8.711.9%), between-run CV (10.5-14.7%), and between-day CV (13.2-21.4%). No HBV DNA was detected in any of 100 samples without HBV, resulting in a clinical specificity of 100%. The levels of HBV DNA showed a good correlation with those determined with Digene Hybrid-Capture II (R2=0.9827). Conclusions : The Real-Q HBV Quantification kit showed a good analytical sensitivity, specificity, and high reliability with a broad reportable range. This assay should be clinically useful in managing patients with HBV infection. (Korean J Lab Med 2006;26:442-8)

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