逆向斑点杂交检测乙型肝炎病毒基因型及YMDD变异

Study of HBV genotypes and YMDD mutations by reverse dot blot

目的应用逆向斑点杂交技术检测乙型肝炎病毒(HBV)基因型及YMDD变异。方法合成有生物素标记的引物,PCR扩增两种DNA片段,与尼龙膜上的探针同时杂交,BCIP/NBT显色。结果检测21例拉米夫啶治疗中的慢性乙型肝炎患者,11例HBV为基因型B,10例为基因型C;YMDD基序有47·6%(10/21)为YMDD,28·6%(6/21)为YVDD,23·8%(5/21)为YIDD。结论在同一张尼龙膜上检测HBV基因型及YMDD变异,经济实惠、准确快捷,且易于开展。

Objective HBV genotypes and YMDD mutations were detected by reverse dot blot. Method Two PCR-amplified fragments generated by biotin labeled primers were complementary to the oligonucleotides that were fixed on the nylon membrane and then detected the signals by BCIP/NBT. Result In 21 Lamivudine therapy patients, there were llgenotype B and 10 genotype C. The YMDD motif was 47. 6% （ 10/21 ） YMDD, 28. 6% （6/21 ） YVDD, and 23.8% （5/21） YIDD. Conclusion Detection of HBV genotypes and YMDD mutations together were economical, exact, rapid and easy.