실시간 정량적 중합효소 연쇄반응을 이용한 혈청 B형 간염 바이러스 DNA 검사 = HBV DNA Quantitation Using Real-time PCR

국문 초록 (Abstract)

배경 및 목적 : 정확한 HBV DNA의 측정은 치료 대상 선정,치료 반응 평가 등에 매우 중요하다. 중합 효소 연쇄 반응법(PCR)을 이용한 HBV DNA 정량법은 검사법의 직선성(linearity)을 나타내는 농도 범위가 좁은 반면, 실시간 정량적 중합 효소 연쇄 반응법(real-time PCR)은 검사 범위가 넓은 장점이 있다. 저자들은real-time PCR로 HBV DNA 정량을 위한 새로운 시발체와 탐촉자를 고안하고, 혈청 HBV DNA치를 측정하여 검출 범위, 반복성 및 재현성을 평가하고 Cobas Amplicor HBV MonitorTM법과 비교하고자 하였다. 대상 및 방법 :HBV DNA에 특이적인 시발체와 탐촉자를 고안하여, 7.0× 104 copies/mL 농도의 EUROHEP standard HBVDNA (NIBSC)를 real-time PCR 정량법의 교정 곡선(calibra-tion curve)과 유효성 평가를 위해 연속 희석하여 사용하였고, 무작위로 추출한 만성 B형 간염 환자 52명의 혈청 HBV DNA치를정량하였다. 정량 결과를 이용하여 검출 범위, 반복성을 평가하고Cobas Amplicor HBV MonitorTM법에 의한 HBV DNA치와 상관성을 평가하였다. 결과 : 본 연구에서 고안한 real-time PCR을 이용한 HBVDNA 정량법의 검출 범위는 6.1× 102 copies/mL부터 6.5× 109copies/mL까지였다. 반복성과 재현성은 CV (coefficient of vari-ation)가 각 3.7-24.9%, 7.8-24.7%였다. 희석하지 않은 본 검사법에 의한 정량 결과와 Cobas Amplicor HBV MonitorTM법에의한 HBV DNA치 비교에서 상관계수 0.8691로 높은상관성을보였다.

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