一种HIV-1耐药基因型检测方法的一致性分析  

Reproducibility evaluation of an in-house HIV-1 drug resistance genotyping test

作　　者：周茜[1,2] 廖玲洁[2] 陈彬[2] 苏俊琪[2] 刘宏伟[3] 王哲[3] 苏斌[4] 董永慧[5] 陈曦[6] 杨绍敏 邵一鸣 邢辉[2] 

机构地区：[1]华中科技大学同济医学院基础医学院病原生物学系,武汉430030 [2]中国疾病预防控制中心性病艾滋病预防控制中心传染病国家重点实验室 [3]河南省疾病预防控制中心 [4]安徽省疾病预防控制中心 [5]新疆维吾尔自治区疾病预防控制中心 [6]湖南省疾病预防控制中心 [7]云南省传染病专科医院艾滋病关爱中心

出　　处：《中国病毒病杂志》2011年第2期110-115,共6页Chinese Journal of Viral Diseases

基　　金：国家"十一五""艾滋病和病毒性肝炎等重大传染病防治"科技重大专项项目(2008ZX10001-004);中国科技部国家重点基础研究发展计划项目(2005CB523103和2005CB522903);中国全球基金艾滋病项目

摘　　要：目的评价一种实验室自建(in-house)HIV-1耐药基因型检测方法的可重复性。方法从2008-2010年已进行HIV-1耐药基因型检测(in-house法)的样品中抽取204份再次进行基因型检测,包括核酸的提取、扩增、测序和序列分析,对两次的耐药结果进行比较。结果 204份样品中,对群体耐药发生的判断上一致性很高,可达98.5%(201/204),在每种抗病毒药物是否发生耐药的判断上[92.2%(188/204)]具有一致性,对每种抗病毒药物的耐药程度判断一致性为81.9%(167/204),84份样品中共有159个位点在两次检测中出现耐药突变位点上不一致的情况,其中不完全一致的位点有149个,蛋白酶区第71位(13/204,6.4%)和逆转录酶区第103位分别出现最多(12/204,5.9%),完全不一致的位点有10个,出现在蛋白酶区第71位,核苷类逆转录酶抑制剂(NRTI)耐药相关突变位点第67、69、70、215、219位,以及与非核苷类逆转录酶抑制剂(NNR-TI)耐药相关突变位点第90、181、221位。结论 in-house方法双重检测对耐药发生及耐药程度的判断上总体具有良好的一致性。但在方法学尤其是混合碱基的判读上需进一步标化以达到更好的可重复性。由于混合碱基的判读会影响耐药突变位点的确定,加强对检测人员的培训,使他们更加严格地执行混合碱基的判断标准和控制序列的质量是非常重要的。Objective To evaluate the reproducibility of an in-house HIV-1 drug resistance（HIVDR） genotyping test.Methods The reproducibility of an in-house HIVDR genotyping test was evaluated with 204 plasma samples,which had been tested from 2008 to 2010.The samples were randomly selected and retested with the same method.A fragment of HIV pol gene was extracted from plasma samples,amplified and sequenced.Drug resistance-related mutations were identified and interpreted through Stanford HIVdb program,and were compared with the results of previous testing.Results The rates of concordance in the overall resistance and resistance to specific antiretroviral drugs were 98.5%（201/204）and 92.2 %（188/204） between the results of the first and second testing,respectively.One hundred and sixty-seven（81.9%）specimens had same levels of resistance to specific antiretroviral drugs between the two sets of results.However,the discordant drug resistance-related mutations were found in 84 samples,among which 159 discordant codons were discovered.Most（149） of the codons were partially discordant.The inconsistent codons were most frequently found at position 71（13/204,6.4%） of the protease and 103（12/204,5.9%） of the reverse transcriptase（RT）, respectively.Ten completely discordant codons were found at position 71 of the protease.Nucleoside reverse transcriptase inhibitor（NRTI） resistance-related sites were found at position 67,69,70,215 and 219 and non-nucleoside reverse transcriptase inhibitor（NNRTI） resistance-related sites were found at position 90,181 and 221 of RT regions,respectively.Conclusions The results revealed high concordance rates in both the incidence and the degree of drug resistance.The reproducibility of the in-house drug genotyping test procedure can be optimized by further standardization and personnel training.

关 键 词：人类免疫缺陷病毒1型 IN-HOUSE 基因型耐药检测 耐药 

分 类 号：R512.91[医药卫生—内科学]