急性乙型肝炎患者血清HBV基因耐药变异及相关因素分析

Analysis of HBV drug-resistant gene mutation and related factors in patients with acute hepatitis B

目的了解急性乙型肝炎(AHB)患者感染HBV基因耐药相关变异性,并对变异相关因素HBV DNA、HBeAg、ALT、AST及TBil进行分析。方法收集AHB患者123例,提取血清HBV DNA对反转录酶(RT)基因区核苷(酸)类似物变异耐药突变10个位点进行检测,同时进行HBV DNA载量、HBeAg、ALT和AST检测。采用焦磷酸测序技术(pyrosequencing)进行DNA测序,数据分析采用PSQ-96MA型焦磷酸测序仪配套的分析软件进行。结果 123例患者中9例发生变异,检出率为7.32%,病毒以变异/未变异共存形式存在。变异位点和变异形式均以204V/I和181V为主,相关耐药主要为拉米夫定、替比夫定和阿德福韦酯。发生变异者与未发生变异者比较,HBV DNA载量、ALT、AST、TBil及HBeAg阴性率差异均无统计学意义(P>0.05)。结论急性HBV感染者体内存在HBV核苷(酸)类似物耐药相关突变,自然变异与HBVDNA载量、ALT、HBeAg、TBil及AST无显著相关性。

Objective To investigate whether the resistant mutants exist in the acute hepatitis B （AHB） patients and variation related factors of HBV DNA, HBeAg, ALT, AST and total bilirubin analysis. Methods Serum were collected from 123 AHB patients after their admission for HBV DNA, drug-resistance variations in HBV reverse transcriptase （RT） gene region nucleoside analogs variation at position of 10 sites were analyzed, while HBV DNA level, HBeAg, ALT and AST were detected. The RT genom region of HBV were amplified by PCR, and the PCR products were sequenced by pyrosequencing. Datas were analyzed using the analysis software PSQ-96MA type pyrosequencing instrument supporting. Results Pyrosequencing showed that drug-resistant variants were positive from 9 cases （7.32%） of 123 AHB patients, and others were not detected mutation, while co-existence of the variants with wild-type virus was common. Variable sites and variant forms were 204V/I and 181V mainly associated with lamivudine, telbivudine and adefovir resistance. The differences of HBV DNA load, ALT, AST, total bilirubin, and HBeAg negative rate were not statistically significant （P〉0.05） between mutants and no-mutants. Conclusion The patients with AHB may be infected by NA-resistant virus, and there was no significant correlation between the natural variation and related factors, such as the level of HBV DNA, ALT, HBeAg, total bilirubin, AST and the status of HBeAg.