바이러스간염 진단을 위한 혈청학적 검사법과 단일 시험관 다중 실시간 중합효소연쇄반응법의 비교 연구

Abstract

배경 : 간염은 여러 가지 원인으로 생길 수 있지만 가장 중요한 원인은 만성 간질환을 유발할 수 있는 간염 바이러스이다. A형 간염은 2004년부터 급격한 증가 추세를 보이고 있고 B형 간염은 여전히 유병률이 높으며 C형 간염은 치명적인 예후를 보인다. 지금까지 간염을 진단하기 위해서 A형 간염바이러스 IgM 항체(IgM anti-HAV), B형 간염 표면항원검사(HBsAg), 항-C형 간염 바이러스 항체(anti-HCV)를 각각 혈청학적 방법으로 검사해야 했으나 분자생물학적 방법으로 세가지 검사를 한번에 시행할 수 있는 단일 시험관 다중 실시간 중합효소연쇄반응법(single tube multiplex real-time polymerase chain reaction)이 개발되어 저자는 그 성능을 평가하고자 하였다. 방법 : 본원에서 IgM anti-HAV, HBsAg, anti-HCV 검사를 동시에 시행한 총 184명의 환자를 후향적으로 조사하였다. IgM anti-HAV, HBsAg, Anti-HCV 측정은 모두 전기화학적발광 면역측정법(electrochemiluminescence immunoassay)을 이용한 Roche Modular Analytics E170 (Roche Diagnostics, Mannheim, Germany)을 사용했으며 기존의 혈청학적 방법과 단일 시험관 다중 실시간 중합효소연쇄반응법의 양성률과 일치율을 분석하였다. 결과 : 총 184개의 검체를 분석한 결과 IgM anti-HAV, HBsAg, anti-HCV의 양성률은 각각 15.2% (28/184), 13.6% (25/184), 8.2% (15/184)로 나타났다. 단일 시험관 다중 실시간 중합효소연쇄반응법으로 검사한 결과 HAV 양성률은 18.5% (34/184), HBV 양성률은 16.3% (30/184), HCV 양성률은 4.3% (8/184)로 나타났고 두 방법간의 일치율은 A형 간염 95.7% (176/184), B형 간염 97.3% (179/184), C형 간염 96.2% (177/184)로 나타났다. 혈청학적 검사에서만 양성으로 판정된 경우는 A형 간염 1예, C형 간염 7예가 있었고 B형 간염은 존재하지 않았다. 단일 시험관 다중 실시간 중합효소연쇄반응법에서만 양성으로 판정된 경우는 A형 간염 7예, B형 간염 5예가 있었으며 C형 간염은 존재하지 않았다. 결론 : 새로이 개발된 단일 시험관 다중 실시간 중합효소연쇄반응법은 기존의 혈청학적 검사법과 95% 이상의 일치율을 보였다. 그러나 C형 간염에 대한 민감도가 낮아 아직까지 혈청학적 방법을 완전히 대체하기는 힘들다고 판단된다. 그럼에도 불구하고 기존의 분자생물학적 검사법의 단점인 검체 처리 및 검사 방법의 어려움이 자동화로 인해 편리해졌으므로 C형 간염바이러스 검출에 대한 민감도만 보완한다면 단일 시험관 다중 실시간 중합효소연쇄반응법을 선별검사로 사용할 수 있을 것이라 생각된다. 간염의 유병률이 높은 우리나라에서 기존의 혈청학적 방법과 일치율이 높으면서 간염바이러스 보균자를 조기에 발견할 수 있고, 세가지 형태의 바이러스를 한번에 검사해 모르고 지나칠 수 있는 중복감염까지 검출하여 심각한 합병증을 사전에 예방할 수 있는 단일 시험관 다중 실시간 중합효소연쇄반응법은 백신과 더불어 간염 유병률을 현격히 감소시킬 수 있는 좋은 검사법이라 생각된다.|Background : There are many causes of human hepatitis. Among them, the most common is viral pathogen. There has been a rapid increase in the prevalence of hepatitis A infection since 2004 and the prevalence of hepatitis B infection is still very high in Korea. Hepatitis B and hepatitis C can lead to serious permanent liver damage, and in many cases, can cause death. To determine the types of hepatitis, each serum pool for the following serological markers need to be determined: hepatitis A IgM antibody (IgM anti-HAV), hepatitis B virus surface antigen (HBsAg), and anti-hepatitis C virus antibody(anti-HCV). Single tube multiplex realtime polymerase chain reaction (STM-RTPCR) simultaneously detects and distinguishes each type of hepatitis A, hepatitis B, and hepatitis C viruses. We investigated the diagnostic performance of STM-RTPCR in comparison with those of serologic tests. Methods : We tested and analyzed 184 serum samples for IgM anti-HAV, HBsAg, and anti-HCV. Serologic markers including IgM anti-HAV, HBsAg, and anti-HCV were performed with the method of electrochemiluminescence immunoassay. We calculated positive and concordance rates of the results between serologic tests and STM-RTPCR. Results : The positive rates of IgM anti-HAV, HBsAg, and anti-HCV using serologic methods were 15.2% (28/184), 13.6% (25/184), and 8.2% (15/184) respectively. The positive rates of the corresponding viral nucleic acid detection by STM-RTPCR were 18.5% (34/184), 16.3% (30/184), and 4.3% (8/184), respectively. The concordance rates between serologic test and STM-RTPCR were 95.7% (176/184) in hepatitis A, 97.3% (179/184) in hepatitis B, and 96.2% (177/184) in hepatitis C. 1 case of hepatitis A and 7 cases of hepatitis C had positive serologic test results despite negative STM-RTPCR results. 7 cases of hepatitis A and 5 cases in hepatitis B had positive STM-RTPCR results despite negative serologies. Conclusion : In our study, the concordance rate between newly developed STM-RTPCR and traditional serologic test is over 95 percent. Since the concordance rate of hepatitis C is still low, the STM-RTPCR cannot replace serologic testing completely. However STM-RTPCR provides a rapid, automated, easy, sensitive and simultaneous detection of three major viruses. As the prevalence rate of hepatitis is high in Korea, despite the low sensitivity of hepatitis C, STM-RTPCR is a useful tool for screening test and early detection of hepatitis viruses. Furthermore we expect that STM-RTPCR contributes to reduce the prevalence of hepatitis in Korea.; Background : There are many causes of human hepatitis. Among them, the most common is viral pathogen. There has been a rapid increase in the prevalence of hepatitis A infection since 2004 and the prevalence of hepatitis B infection is still very high in Korea. Hepatitis B and hepatitis C can lead to serious permanent liver damage, and in many cases, can cause death. To determine the types of hepatitis, each serum pool for the following serological markers need to be determined: hepatitis A IgM antibody (IgM anti-HAV), hepatitis B virus surface antigen (HBsAg), and anti-hepatitis C virus antibody(anti-HCV). Single tube multiplex realtime polymerase chain reaction (STM-RTPCR) simultaneously detects and distinguishes each type of hepatitis A, hepatitis B, and hepatitis C viruses. We investigated the diagnostic performance of STM-RTPCR in comparison with those of serologic tests. Methods : We tested and analyzed 184 serum samples for IgM anti-HAV, HBsAg, and anti-HCV. Serologic markers including IgM anti-HAV, HBsAg, and anti-HCV were performed with the method of electrochemiluminescence immunoassay. We calculated positive and concordance rates of the results between serologic tests and STM-RTPCR. Results : The positive rates of IgM anti-HAV, HBsAg, and anti-HCV using serologic methods were 15.2% (28/184), 13.6% (25/184), and 8.2% (15/184) respectively. The positive rates of the corresponding viral nucleic acid detection by STM-RTPCR were 18.5% (34/184), 16.3% (30/184), and 4.3% (8/184), respectively. The concordance rates between serologic test and STM-RTPCR were 95.7% (176/184) in hepatitis A, 97.3% (179/184) in hepatitis B, and 96.2% (177/184) in hepatitis C. 1 case of hepatitis A and 7 cases of hepatitis C had positive serologic test results despite negative STM-RTPCR results. 7 cases of hepatitis A and 5 cases in hepatitis B had positive STM-RTPCR results despite negative serologies. Conclusion : In our study, the concordance rate between newly developed STM-RTPCR and traditional serologic test is over 95 percent. Since the concordance rate of hepatitis C is still low, the STM-RTPCR cannot replace serologic testing completely. However STM-RTPCR provides a rapid, automated, easy, sensitive and simultaneous detection of three major viruses. As the prevalence rate of hepatitis is high in Korea, despite the low sensitivity of hepatitis C, STM-RTPCR is a useful tool for screening test and early detection of hepatitis viruses. Furthermore we expect that STM-RTPCR contributes to reduce the prevalence of hepatitis in Korea.