血清乙型肝炎病毒RNA定量分析系统的建立
肝炎病毒,,肝炎病毒，乙型；转录体；癌，肝细胞；血清；聚合酶链反应；拉米夫定;,疗效评定,0引言,1材料和方法,2结果,3讨论,【参考文献】
(第四军医大学唐都医院病理科，陕西 西安 710038，Division of VirusHost Interactions, German Cancer Research Center, Heidelberg, Germany, 天津医科大学肿瘤医院，天津 300060，陕西省肿瘤医院, 陕西 西安 710061)Quantitative assay system established for different hepatitis B virus RNAs in sera sample
ZHANG Wei, SU Qin, LIU Jie, Hans J. Hacker, NIU Yun, LIANG XiuFen, Claus H. Schroeder
Department of Pathology, Tangdu Hospital, Fourth Military Medical University, Xian 710038, China, Division of VirusHost Interactions, German Cancer Research Center, Heidelberg, Germany, Tianjin Cancer Hospital, Tianjin Medical University, Tianjin 300060, China, Shaanxi Cancer Hospital, Xian 710061, China
【Abstract】AIM： To establish a quantitative system to characterize different RNA molecules (transcripts) of hepatitis B virus (HBV) in circulation. METHODS： Viral nucleic acids were extracted from serum samples and HBV DNA and RNA were characterized quantitatively by competitive PCR and RT-PCR procedures. RESULTS： A seroassay was established to characterize 3′-end structure of the full-length and truncated viral transcripts. Copy numbers of viral RNA/DNA segments corresponding to X, Core and XPrecore regions were determined, representing early, middle and late stages of HBV gene replication, respectively. In addition, the data demonstrate dynamic changes in the circulating viral transcripts as well as DNA in their copy numbers and structures during lamivudine therapy. After a 8-week treatment, the copy numbers for C or PreC/X DNA segments decreased to 105·mL-1 from 109·mL-1, significantly below that for X segment (from 109·mL-1 to 107·mL-1). There was no significant decrease in RNA copy numbers. Polyadenylated HBV RNA was also determined using anchored oligo (dT) primers targeting fRNA and trRNA. The copy numbers of fRNA and trRNA were 105 copies per mL of serum during most of the treatment period, significantly below that of X segment (107·mL-1). The excess of X segment RNA over fRNA levels suggested a packagingrelated removal of poly (A) 3′ends. CONCLUSION： An assay for the detection of copy numbers and different 3′end structures of the circulating HBV transcripts is established, which provides a more precise approach to the detection of dynamic change of circulating HBV transcripts. ......
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