中国人终末期肝病组织中乙肝病毒转录体的检测

Detection of hepatitis B virus transcript in tissues of end-stage liver disease in Chinese

目的 :探讨肝组织中HBVDNA和病毒转录体与终末期肝病的关系 .方法 :用TRIzol从肝组织中提取组织核酸 ,PCR及RT PCR扩增HBVXDNA ,XRNA ,fRNA和trRNA ,琼脂糖凝胶电泳显示产物 ,Southern blot杂交验证PCR扩增的可靠性 .结果 :2 7例HBsAg(+)肝组织 ,1 9例HBxDNA/RNA检测均为阳性 ,其中HBeAg(+)者fRNA检出率较HBeAg(- )者高 ;2 2例HBsAg(- )肝组织 ,有 1 6例可检测到HBxDNA或RNA ,HBsAg(- )的隐匿性感染在中国人终末期肝病中有较高的检出率 72 .7% ;HBV病毒转录体trRNA在肝癌及肝硬化组织中的检出率明显高于其他肝脏疾病 (P <0 .0 5 ) .结论 :HBV隐匿性感染与中国人终末期肝病的发生密切相关 。

AIM: To explore the relationship between the hepatitis B virus DNA as well as transcript in liver tissues and end stage liver disease. MEHTODS: Nucleic acid was distilled with TRIzol reagent from liver tissues. Hepatitis B virus (HBV) X region deoxyribonucleic acid (DNA), ribonucleic acid (RNA), full length RNA (fRNA) and truncated RNA (trRNA) were amplified by polymerase chain reaction (PCR) and reverse transcriptase polymerase chain reaction (RT PCR), and their products were displayed by agarose electrophoresis. The dependability of PCR was validated by Southern blot. RESULTS: Positive HBx DNA and RNA were found in 19 cases of the 27 HbsAg (+) liver tissues and positive fRNA in HbeAg (+) liver tissues was higher than that in HbeAg (-) liver tissues. Positive HBx DNA or RNA was found in 16 cases of the 22 HbsAg (-) liver tissues and the HbsAg (-) occult infection had a high positive result (72.7%) in end stage liver disease in Chinese. Positive HBV transcript trRNA was significantly higher in liver cancer and hepatocirrhosis than that in other liver diseases ( P <0.05). CONCLUSION: The occult infection of HBV and HBV trRNA in liver tissues is closely related to the occurrence of end stage liver disease in Chinese.