慢性HBV感染者恩替卡韦耐药突变特征分析  

Hepatitis B virus （HBV） gene mutations related to entecavir-resistance in patients with chronic HBV infections

作　　者：高国生[1] 翁彭剑[1] 许小敏[1] 董飞波 颜华东[2] 胡爱荣[2] 

机构地区：[1]浙江省宁波市第二医院检验科,315010 [2]浙江省宁波市第二医院肝病科

出　　处：《中华临床感染病杂志》2015年第2期139-142,共4页Chinese Journal of Clinical Infectious Diseases

基　　金：2014年浙江省医药卫生省部培育计划（2014PYA018）;宁波市自然科学基金（2013A610239）

摘　　要：目的分析慢性乙型肝炎病毒（HBV）感染者恩替卡韦（ETV）耐药突变特征。方法采用实时荧光定量聚合酶链反应（PCR）方法对2010年2月至2014年5月宁波市第二医院44例ETV耐药的慢性HBV感染者的血清HBV多聚酶区进行扩增，并对PCR产物进行直接测序。采用SPSS16．0软件分析ETV耐药相关位点的突变发生率，及其与基因型及临床特征的相关性。结果44例患者的基因突变模式以rtS202G／I（52．28％，23／44）为主，其次为rtT184A／G／I／S（36．36％，16／44）和rtM250V／L（11．36％，5／44）。共检出9种基因突变模式，其中以rtL180＋rtM204V＋rtS202G／I和rtL180＋rtM204V＋rtT184A／G／I／S最为常见，分别占38．64％（17／44）和27．27％（12／44）。基因B型和C型患者ETV耐药突变位点分布差异有统计学意义（χ^2=12．294，P〈0．01）。比较不同基因突变模式感染患者的临床资料发现，发生rtT184A／G／I／S突变的患者肝脏损害程度较其他患者更为严重（χ^2=14．499，P〈0．01），而发生rtM250V／L突变的患者HBeAg阳性率较其他模式低（χ^2=10．057，P〈0．01）。结论对ETV耐药的慢性HBV感染者血清HBV多聚酶区基因突变以rtL180＋rtM204V＋rtS202G／I较为常见，且不同基因突变模式与基因分型、肝脏损害程度和HBeAg阳性率可能相关。Objective To investigate hepatitis B virus （HBV） gene mutations related to entecavir （ETV）-resistance in patients with chronic HBV infections. Methods Serum samples were collected from 44 patients with chronic HBV infections and resistant to ETV treatment who were admitted in Ningbo No. 2 Hospital during February 2010 and May 2014. The HBV polymerase regions were amplified by real-time fluorescent quantitative polymerase chain reaction （PCR） method, and the PCR products were analyzed with direct sequencing. SPSS 16.0 was used to assess the frequency of HBV polymerase gene mutations, and its relation to the viral genotype and clinical features. Results The most common HBV polymerase gene mutation was rtS202G/I （ 52. 28%, 23/44 ）, followed by rtT184A/G/I/S （ 36. 36%, 16/44 ） and rtM250V/L （11. 36%, 5/44）. Nine mutation patterns were deteeted, in which rtL180 ＋ rtM204V ＋ rtS202G/I （38.64%, 17/44） and rtL180 ＋ rtM204V ＋ rtT184A/G/I/S （27.27%, 12/44） were the most frequent ones. The difference in gene mutations between genotype B and C was of statistical significance （χ^2=12.294, P〈0.01 ）. Patients carrying rtT184A/G/I/S mutations were associated with worse liver function （χ^2 = 14. 499, P 〈 0. 01 ） , and those carrying rtM250V/L mutations were associated with lower HBeAg positive rate （χ^2 = 10.057, P〈0. 01）. Conelusions rtL180M ＋rtM204V ＋rtS202G/I is the most common HBV polymerase gene mutation related to ETV resistance in patients with chronic HBV infections. Different gene mutations may be associated with HBV genotypes, severity of liver damages, and HBeAg positive rate.

关 键 词：肝炎 乙型 慢性 恩替卡韦 抗药性 DNA突变分析 

分 类 号：R512.62[医药卫生—内科学]