PCR-反向点杂交法检测 HBV 耐药变异与基因型的探讨

摘要

目的：探讨 PCR-反向点杂交法检测 HBV 耐药变异与基因型的相关性及 HBV 变异位点与肝功能指标、HBVDNA病毒载量的关系。方法筛选符合条件的462例服用核苷类药物治疗的慢性乙型肝炎患者血清标本，用 PCR-反向点杂交法检测 HBV 的耐药突变位点以及基因型，并对 HBV 耐药变异与基因型、肝功能检测指标、HBV DNA 病毒载量等指标进行相关性分析。结果在462例服用核苷类药物治疗的慢性乙型肝炎患者血清中检测耐药变异株45例，变异率约为9．74％；其中180M 和204I/V 突变株16例，35．5％；204V 突变株6例，13．3％；204I 突变株13例，28．9％；180V 突变株3例，6．7％；181V 突变株3例，6．7％；207I 突变株1例，2．2％，236T 突变株3例，6．7％。HBV 基因型中 B 基因型105例，C 基因型337例，B 基因型和 C 基因型联合感染4例，D 基因型0例，其他基因型2例。将检测结果与临床资料进行相关性分析：乙肝患者的耐药变异位点与基因型、肝功能指标 ALT 无相关性（P ＞0．05），与 HBVDNA 病毒载量有一定相关性（P ＜0．05）。结论1．茂名地区的 HBV 基因型可能与其他南方地区的基因型有差异，以 HBV-C 基因型为主；2．PCR-反向点杂交法是一种快速、准确发现核苷类药物治疗后耐药位点的检测方法；3．通过临床资料分析显示乙肝患者的耐药变异位点与肝功能指标 ALT 间差异无统计学意义（P ＞0．05），即无相关性，HBV-DNA 病毒载量间差异有统计学意义（P ＜0．05），即有一定相关性。%Objective To study the correlation between the drug-resistance variation and the genotypes of hepatitis B virus (HBV)detected by the PCR-reverse dot blot and the relation between the HBV variation loci with the liver function indexes and HBV DNA viral loading.Methods The serum samples from 462 patients with chronic hepatitis B treated by oral nucleoside drugs were screened.The PCR-reverse dot blot was adopted to detect the drug-resistance gene mutation loci and genotypes.The correla-tion between the HBV drug-resistance mutant with the genotypes,liver function indexes and HBV DNA viral loads was performed. Results Among 462 patients taking nucleoside drugs for treating chronic hepatitis B,45 drug-resistance mutants were detected with the mutation rate of 9.74%;in which,16 cases (35.5%)were 180M and 204I/V mutant,6 cases(13.3%)were 204V,13 ca-ses(28.9%)were 204I mutant,3 cases (6.7%)were 180V mutant and 3 cases(6.7%)were 236T mutant.The HBV genotyping showed 105 cases of genotype B,337 cases of genotype C,0 case of genotype D and 2 cases of other genotypes.Conclusion (1)The HBVgenotypes in Maoming area may be different from the genotypes in other southern regions and is dominated by HBV-C geno-type.(2)The PCR-reverse dot blot method is a detection method for fastly and accurately finding the drug-resistance loci after nu-cleosides therapy.(3)The clinical analysis demonstrates that the drug-resistance mutation loci has no correlation with the liver func-tion index ALT(P >0.05),but there was certain correlation between the drug-resistance mutation loci in hepatitis B and HBV DNA viral load(P <0.05).