Detection of hepatitis B virus genome in hepatocellular carcinoma tissues with PCR-in situ hybridization
Detection of hepatitis B virus genome in hepatocellular carcinoma tissues with PCR-in situ hybridization
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DOI：
10.1016/s0166-0934(99)00040-3
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发表时间：
1999-07-01
影响因子：
3.1
通讯作者：
Li, C
中科院分区：
文献类型：
--
作者：
Shieh, B;Lee, SE;Li, C
本文报道了14例B肝炎病毒（HBV）血清阳性患者肝细胞癌组织标本中HBV DNA的检测。检测是通过聚合酶链反应（PCR）扩增的目标序列，然后通过特异性定位的PCR产物与原位杂交。FISH（PCR-原位杂交）在大多数肿瘤组织中产生强阳性信号，尽管染色体整合的HBV基因组拷贝数非常低，而在对照样品中未检测到信号，表明信号是HBV特异性的。有时在肿瘤周围的结节中检测到阳性信号，表明HBV感染了未转化的肝细胞。HBV-DNA在细胞核和细胞质中检测到一些标本，可能代表HBV在生命周期的不同阶段。在一种情况下，显示了病毒DNA的梯度，最高的DNA信号集中在病毒抗原表达的位点。综上所述，FISH被证明是一种高灵敏度的分子检测方法，能够原位检测低拷贝数病毒基因组的存在。(C)1999 Elsevier Science B. V.保留所有权利。
The detection is described of hepatitis B virus (HBV)-DNA in preserved hepatocellular carcinoma tissues, which were derived from 14 HBV-seropositive patients. Detection was by polymerase chain reaction (PCR) amplification of the target sequence, followed by specific localization of the PCR product with in situ hybridization. FISH (PCR-in situ hybridization) yielded strong positive signals in most of the tumor tissues despite very low copy numbers of chromosome-integrated HBV genome, whereas no signal was detected in control samples, indicating that the signals were specific for HBV. Positive signals were sometimes detected in cirrhotic nodules surrounding the tumor regions, indicating that HBV had infected non-transformed liver cells. HBV-DNA was detected in both nucleus and cytoplasm in some specimens, possibly representing HBV at different stages of the life cycle. In one case, a gradient of viral DNA was revealed, with the highest DNA signal centered at the site of viral antigen expression. Taken together, FISH is shown to be a highly sensitive molecular detection method that is capable of detecting the presence of a low copy number viral genome in situ. (C) 1999 Elsevier Science B.V. All rights reserved.