HBVP区聚合酶基因变异研究

Detection and Analysis of Gene Mutation of HBV DNA

目的 建立快速特异的HBV基因变异检测技术及变异株的酶切位点分析。方法 对酪氨酸、缬氨酸、天冬氨酸、天冬氨酸 (YVDD)及酪氨酸、异亮氨酸、天冬氨酸、天冬氨酸 (YIDD)变异病人进行测序分析。结果 在HBV感染的病例中 ,不仅检出酪氨酸、蛋氨酸、天冬氨酸、天冬氨酸 (YMDD)到YIDD /YVDD变异 ,还存在亮氨酸、亮氨酸、亮氨酸、亮氨酸 (LLLL)到亮氨酸、蛋氨酸、亮氨、亮氨酸 (LMLL)变异 ,天冬氨酸、亮氨酸、组氨酸、天冬氨酸 (DLHD)到天冬氨酸、蛋氨酸、组氨酸、天冬氨酸 (DMHD)变异及亮氨酸、亮氨酸、丙氨酸、谷氨酸 (LLAQ )到亮氨酸、蛋氨酸、丙氨酸、谷氨酸 (LMAQ)变异。酶切位点分析结果表明不同的变异株存在不同的限制性酶切位点。结论 提示这些变异位点均在“ATG”(M)蛋氨酸密码子尚需进一步探讨研究。YVDD、YIDD。

Objective To develop a rapid specific method to detect HBV gene mutation by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Methords The patients of YMDD and YIDD mutation were detected by DNA sequencing.Results the DNA sequencing demonstrated that not only YMDD and YIDD/YVDD mutation, but LLLL,LMLL,DLHD,DMHD,LLAQ and LMAQ mutation were also detected in different hepatitis B patients. The results of PCR-RFLP indicated that there were different restricted sites in the distinct mutation.Conclusion The results above indicated that all these mutants exist in the initiator codon of ATG(Met). The mutation of LMLL DMHD LMAQ was associated with Lamivordin treatment. Whether mutation of Met links with the second replicating of HBV needs to be further studied.