乙型肝炎病毒不同基因型 DNA聚合酶对病毒复制水平的影响

Influence of HBV polymerase on virus replication with different genotypes

摘要:

目的：研究不同基因型聚合酶（polymerase，pol）复制乙型肝炎病毒（hepatitis B virus，HBV）DNA的能力是否存在差异。方法：采用不依赖酶切和连接的克隆（restriction digestion and ligation-independent cloning，RLIC）方法和HBV聚合酶反式互补技术表达并检测A、B、C、D 4种基因型聚合酶复制HBV DNA的情况。结果：（1）反式互补技术可使 A、B、C、D型 HBV 重新获得表达，HBV DNA量分别为A=（63.03±8.03）×105 copies/μl，B=（23.23±3.53）×105 copies/μl，C=（17.63±1.25）×105 copies/μl，D=（75.73±9.43）×105 copies/μl。（2）HBV不同基因型聚合酶之间复制HBV能力存在差异，F（4，10）=91.88，P＝0.000。（3）A型和D型聚合酶复制HBV能力强于B型和C型聚合酶，差异有统计学意义，P（A，B）=0.001，P（A，C）=0.00１，P（B，D）=0.00１，P（C，D）=0.00１。结论：HBV不同基因型聚合酶之间复制HBV能力存在差异；为深入研究不同基因型在地理分布、所致疾病严重性等方面提供了有用信息并奠定了坚实的基础。

Abstract:

Objective：To investigate hepatitis B virus（ＨＢＶ） DNA levels with polymerase of HBV genotypes A，B，C and D. Methods：The method of restriction digestion and ligation-independent cloning（RLIC） and trans-complementation by HBV polymerase was used to express and test HBV DNA levels with four genotypes. Results：（1）HBV expressed again after HBV polymerase trans-complementa－tion. HBV DNA：A=（63.03±8.03）×105 copies/μl，B=（23.23±3.53）×105 copies/μl，C=（17.63±1.25）×105 copies/μl，D=（75.73±9.43）×105 copies/μl. （2）There were differences in HBV DNA levels among different genotype polymerases，F（4，10）=91.88，P＝0.000. （3）HBV DNA levels were higher in genotype D and A than in genotype B and C，P（A，B）=0.001，P（A，C）=0.00１，P（B，D）=0.00１，P（C，D）=0.00１. Conclusion：HBV DNA levels with polymerase of hepatitis B virus genotypes A，B，C and D were different；the mechanism of differ－ences remains to be further studied.