Lampertico P, Malter J S, Colombo M, Gerber M A
Institute of Internal Medicine, University of Milan, Italy.
Am J Pathol. 1990 Aug;137(2):253-8.
DNA isolated from formalin-fixed, paraffin-embedded liver tissues from nine patients with hepatocellular carcinoma and six control patients was screened for hepatitis B virus (HBV) DNA with surface (S) and core (C) gene-specific primers by a modification of the polymerase chain reaction--southern blot technique (PCR-SB). PCR-SB results were correlated with histologic, immunohistochemical, and serologic findings. All cases with an established HBV etiology were positive by PCR-SB, as were three cases with negative immunohistochemistry and serology. Often there was selective amplification with one primer set and, in two cases, smaller than expected HBV amplification products suggesting internal deletions. The presence of a potent PCR inhibitor in nucleic acid preparations from tissue blocks that can be removed by Sephadex G-50 chromatography was confirmed. PCR-SB will be a powerful method for the diagnosis and follow-up of patients with HBV infection and may provide new insights into viral hepatocarcinogenesis.
采用改良的聚合酶链反应- Southern印迹技术(PCR-SB),使用表面(S)和核心(C)基因特异性引物,对9例肝细胞癌患者和6例对照患者的福尔马林固定、石蜡包埋肝组织中分离的DNA进行乙型肝炎病毒(HBV)DNA筛查。PCR-SB结果与组织学、免疫组织化学和血清学结果相关。所有已确定为HBV病因的病例经PCR-SB检测均为阳性,3例免疫组织化学和血清学检测阴性的病例也是如此。经常会出现一组引物的选择性扩增,在两例病例中,HBV扩增产物小于预期,提示存在内部缺失。证实了组织块核酸制剂中存在一种有效的PCR抑制剂,可通过Sephadex G-50柱色谱法去除。PCR-SB将成为诊断和随访HBV感染患者的有力方法,并可能为病毒性肝癌发生提供新的见解。