Detection of integrated hepatitis B virus DNA in hepatocellular carcinoma cell lines by nonradioactive in situ hybridization
Detection of integrated hepatitis B virus DNA in hepatocellular carcinoma cell lines by nonradioactive in situ hybridization
复制标题
非放射性原位杂交检测肝癌细胞系中整合的乙型肝炎病毒DNA
作者：
Nicholas Tay;S. Chan;E. Ren
建立了一种灵敏、特异的非放射性原位杂交方法，用于检测肝细胞癌（HCC）细胞系中单拷贝整合的B型肝炎病毒（HBV）DNA序列。在96孔微量滴定板中进行生物素化HBV（adr，adw）DNA探针与9种不同人HCC细胞系的原位杂交。在HCC细胞系HCCM、Hep 3B、huH-1、huH-4和PLC/PRF/5中检测到整合。在Hep 3B和huH-4中也实现了单拷贝HBV DNA序列的检测。HCC细胞系HepG 2、HUH-6、HUH-7、Mahlavu和非HCC对照MCF-7给出了明确的阴性结果。这些结果与Southern印迹杂交法所得结果具有100%的相关性。结果表明，通过所述方法可以非放射性检测HCC细胞系中单拷贝整合的HBV DNA序列，这对于需要原位杂交的病毒基因组分析具有潜在的应用价值。
A sensitive and specific nonradioactive in situ hybridization method capable of detecting single‐copy integrated hepatitis B virus (HBV) DNA sequences in hepatocellular carcinoma (HCC) cell lines was developed. In situ hybridization of biotinylated HBV (adr, adw) DNA probes with nine different human HCC cell lines were carried out in 96‐well microtiter plates. Integration was detected in HCC cell lines HCCM, Hep3B, huH‐1, huH‐4, and PLC/PRF/5. Detection of single‐copy HBV DNA sequences was also achieved in Hep3B and huH‐4. HCC cell lines HepG2, HUH‐6, HUH‐7, Mahlavu, and the non‐HCC control MCF‐7, gave clear negative results. These results show a 100% correlation with those obtained by Southern blot hybridization assay. The results demonstrate that nonradioactive detection of single‐copy integrated HBV DNA sequences in HCC cell lines is possible by the method described, which has potential application for viral genome analysis requiring in situ hybridization.
影响因子：
158.5
作者：
BRECHOT, C;DEGOS, F;BERTHELOT, P
通讯作者：
BERTHELOT, P
DOI：
10.1073/pnas.80.13.4045
发表时间：
1983-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
LEARY, JJ;BRIGATI, DJ;WARD, DC
通讯作者：
WARD, DC