Rijntjes P J, Van Ditzhuijsen T J, Van Loon A M, Van Haelst U J, Bronkhorst F B, Yap S H
Am J Pathol. 1985 Sep;120(3):411-8.
To examine the relationship of hepatitis B virus (HBV) DNA sequences in the liver with histopathologic features and antigenic markers, the authors determined the hepatocytic status of viral DNA by in situ hybridization in formalin-fixed liver sections using a biotinylated probe in 45 patients with various chronic liver diseases. The results were compared retrospectively with the HBV serologic markers and histopathologic features including the presence of ground-glass cells or Shikata staining positivity. The specificity of this in situ detection of HBV DNA has been proven excellent in a double-blind control study in 18 patients in whom liver HBV DNA was also determined by DNA extraction, gel electrophoresis, and the Southern blotting technique. In 41 patients, the findings of HBV DNA and serologic markers were concordant (17 positive and 24 negative). Twelve of the 20 HBV-DNA-positive patients were HBsAg-positive (6 with chronic hepatitis, 3 with cirrhosis, and 3 with hepatocellular carcinoma). Ground-glass cells or Shikata positivity were found in 10 of these 12 patients. HBV DNA sequences were found in the liver of all patients with chronic liver disease and serologic positivity for HBV infection. In liver with normal histologic features, HBV DNA was not demonstrable, despite the positive anti-HBc and anti-HBs. However, a positive HBV DNA was found in 3 serologically negative patients. In another patient the interpretation of findings was impossible because of severe hemosiderosis. From this study, it is concluded that in situ detection of HBV DNA in formalin-fixed liver sections has a clinical value and is suitable for routine use.
为了研究肝脏中乙肝病毒(HBV)DNA序列与组织病理学特征及抗原标志物之间的关系,作者采用生物素化探针,通过原位杂交技术,对45例患有各种慢性肝病的患者的福尔马林固定肝脏切片进行检测,以确定病毒DNA的肝细胞状态。将结果与HBV血清学标志物及组织病理学特征进行回顾性比较,这些特征包括毛玻璃样细胞的存在或Shikata染色阳性。在一项双盲对照研究中,对18例患者进行了检测,其中通过DNA提取、凝胶电泳和Southern印迹技术也测定了肝脏HBV DNA,结果证明这种原位检测HBV DNA的特异性非常好。在41例患者中,HBV DNA和血清学标志物的检测结果一致(17例阳性,24例阴性)。20例HBV-DNA阳性患者中有12例HBsAg阳性(6例为慢性肝炎,3例为肝硬化,3例为肝细胞癌)。这12例患者中有10例发现了毛玻璃样细胞或Shikata染色阳性。在所有慢性肝病且HBV感染血清学阳性的患者肝脏中均发现了HBV DNA序列。在组织学特征正常的肝脏中,尽管抗-HBc和抗-HBs阳性,但未检测到HBV DNA。然而,在3例血清学阴性患者中发现了HBV DNA阳性。在另一例患者中,由于严重的含铁血黄素沉着症,无法对结果进行解读。从这项研究可以得出结论,在福尔马林固定肝脏切片中进行HBV DNA的原位检测具有临床价值,适用于常规使用。