HBV携带者血清病毒标志物和HBV DNA与肝组织中HBVcccDNA相关性的研究

Quantitative analysis of HBV cccDNA from liver biopsy specimens in HBV carriers： correlation with serological makers and sera HBV DNA

目的探讨HBV携带者血清病毒标志物和HBV DNA与肝组织中HBVcccDNA之间的关系。方法应用实时荧光定量聚合酶链反应（RT-PCR）方法检测30例经肝组织活检病理检查确定为HBV携带者的肝组织中HBVcccDNA、HBVtDNA和血清HBV DNA，同时用化学发光免疫分析法检测HBsAg、HBeAg定量，分析感染者肝组织内HBVcccDNA与肝组织内HBVtDNA、血清HBVDNA、HBsAg及HBeAg定量水平之间的关系。结果HBV携带者肝组织中均可检出HBVcccDNA，范围在3．15×10^3拷贝／mg～1．06×10^7拷贝／mg（对数值：5．66±O．93）；肝组织cccDNA定量与肝组织总HBVDNA定量呈正相关（r=0．375，P〈0．05），与血清HBVDNA无相关性（r=0．174，P〉0．05）。肝组织中HBVcccDNA水平与血清HBsAg定量呈高度正相关（r=0．562，P〈0．001）；而与血清HBeAg定量无相关性（r=0．152，P〉0．05）。结论HBV携带者肝组织内HBVcccDNA成稳定的中等水平复制；血清HBVDNA载量不能直接代表其肝组织中的HBVcccDNA水平；血清HBsAg定量可作为反映肝幺日织中HBVcccDNA水平的指标。

Objective To investigate the correlation of sera HBV DNA and serological makers with hepatic tissue HBVcccDNA in chronic HBV carriers. Methods Real time fluorescence quantitative polymerase chain reaction （RT-PCR） were used to detect HBV covalently closed circular DNA （cccDNA） and total intrahepatic HBV DNA from 30 needle-biopsy specimens as well as HBV DNA in sera in chronic HBV carriers. Quantification of the HBsAg, HBeAg in sera were quantified using Chemiluminescence immunoassay. Results HBVcccDNA can be detected in chronic HBV carriers, which rang from 3.15 ±10^3 copies/mg to 1.06 ±10^7 copies/mg. There was a positive correlation between the cccDNA and HBVtDNA （r=0. 375, P 〈0.05）, but there was no correlation between the cccDNA and sera HBV DNA （P = 0. 174）. There was a positive correlation between eccDNA and sera HBsAg quantification （r =0. 562, P 〈 0. 001 ） but no correlation with sera HBeAg qantification （ r = 0. 152, P 〉 O. 05 ）. Conclusion HBV cccDNA can be replicated stably in hepatic tissue in all chronic HBV carriers. HBV DNA in sera can not be indicated hepatic tissue cecDNA level. While HBsAg quantification in sera can be used as a marker of eccDNA quantification in hepatic tissue to some extent.