摘要
目的:建立以PCR-RFLP检测HBV多聚酶M204V/M204I和L180M变异的方法,探寻上述位点变异之间的关系。方法:构建质粒pUCm-V+M/I+L,采用基因序列测定技术以确证PCR-RFLP检测HBV多聚酶M204V/M204I和L180M变异的方法。采用建立的方法对拉米夫定治疗过程中发生耐药变异的41例慢性HBV感染患者进行YMDD变异类型的检测。结果:①建立了检测HBV多聚酶M204V/M204I和L180M变异的PCR-RFLP方法,且与基因序列测定的结果一致;②HBV多聚酶M204V变异,占YMDD变异的31.7%(13/41),均伴有L180M变异;M204I变异占YMDD变异的68.3%(28/41),其中46.4%(13/28)伴有L180M变异。结论:①HBV多聚酶M204V/M204I和L180M变异的PCR-RFLP检测方法可靠;②M204V变异均伴有L180M变异,46.4%的M204I变异也伴有L180M变异。
Objective: To detect M204V/M204I and L180M mutations in hepatitis B virus DNA polymerase associated with lamivudine-resistance by PCR-RFLP. Methods: A PCR-RFLP assay was set up to detect M204V/M204I and L180M mutations in hepatitis B virus DNA polymerase, and plasmid pUCm-V+M /I+L was constructed as control for these mutations. Both of the assays were identified by HBV polymerase gene sequencing. Forty-one samples from patients with chronic hepatitis B and lamivudine-resistance were determined using PCR-RFLP. Results: ①A PCR-RFLP assay was established and confirmed to be useful for detecting M204I/M204V and L180M mutations in HBV DNA polymerase. M204V mutations occurred in 31.7% (13/41) of YMDD mutations, all accompanied by L180M mutations. M204I mutations developed in 68.3% (28/41), 46.4% (13/28) of them accompanied by L180M mutations. Conclusion: ①A PCR-RFLP assay is reliable to determine M204V/M204I and L180M mutations in hepatitis B virus DNA polymerase; all M204V mutations are always accompanied by L180M mutations, but about half of M204I are also accompanied by L180M mutations.
出处
《山东大学学报（医学版）》
CAS
北大核心
2005年第2期145-148,共4页
Journal of Shandong University:Health Sciences
基金
山东省卫生厅计划项目(2001CA1CAA11)
首都医学发展科研基金项目(2002-3046)。
关键词
拉米夫定
肝炎病毒
乙型
抗药性
Lamivudine
Hepatitis B virus
Drug resistance
