对in—houseHIV-1基因型耐药性检测方法的评价  

Evaluation of an in-house method for HIV-1 drug resistance genotyping test

作　　者：耿庆茂[1] 李韩平[1] 辛天义 庄道民[1] 鲍作义[1] 刘永健[1] 李林[1] 王铮[1] 刘思扬[1] 李敬云[1] 

机构地区：[1]全军艾滋病检测中心,军事医学科学院微生物流行病研究所,病原微生物安全国家重点实验室,北京100071 [2]河南省沈丘县卫生局

出　　处：《中华检验医学杂志》2011年第9期849-854,共6页Chinese Journal of Laboratory Medicine

基　　金：国家自然科学基金资助项目（30830088、30800938）

摘　　要：目的评价in—houseHIV—1基因型耐药检测方法的敏感性与准确性。方法收集2004年4月至2008年10月全军艾滋病检测中心检测的来自河南、广西130份血浆标本。以美国FDA批准的HIV-1基因型耐药性检测系统（ViroSeq^TMv2．0）为参考方法，并与建立起的基因型耐药性检测方法（in—house）平行检测待检样本，比较二者在扩增效率、耐药突变位点检测以及耐药报告等方面的一致性。结果已知的14850个耐药突变位点中，2种方法可同时对99．3％（14752／14850）的耐药突变位点准确检出；在对不同突变位点的检测中，2种方法对蛋白酶抑制剂耐药突变位点、逆转录酶抑制剂耐药突变位点及两类抑制剂耐药突变位点检测一致率分别为99．7％、99．0％和99．3％（Kappa值分别为0．9099、0．9521和0．9488，P值均〈0．01）；2种方法出具的两类药物的耐药结果报告一致率94．6％（Kappa=0．6374，P〈0．01）。本研究共检测到34个ViroSeq“数据库（ViroSeq^TMsoftwarev2．7）未收录位点，其中2个突变位点对耐药的影响较大。结论in—house基因型耐药性检测方法与ViroSeq^TM基因型耐药性检测系统在耐药突变位点检测以及评价上具有高度一致性，是一种快速准确、性价比高的HIV-1基因型耐药检测方法，同时在耐药数据库方面具有一定优势。Objective To evaluate the sensitivity and accuracy of an in-house detecting method of HIV-1 genotypic drug resistance system. Methods Totally 130 serum specimens from Henan and Guangxi province were collected from April 2004 to October 2008 and tested in the Military HIV Testing Center of China. ViroSeqTM v2.0 （Abbott, Switzerland） , a US FDA approved HIV genotypic drug resistance detecting system was utilized as the reference method. All the specimens were detected by the novel in-house method and the reference method to validate the difference in amplifying efficiency, drug resistance mutation detection and drug resistance report. Results Concerning the 14 850 known drug resistance mutation sites, 14 752 （99. 3% ） mutations can be detected by both of the two methods. Rates of concordance of detection in the regions of protease inhibitors-, reverse transcriptase inhibitors- and both two classes inhibitors-resistance were 99.7% （ Kappa = 0. 909 9, P 〈 0. 01 ） , 99. 0% （ Kappa = 0. 952 1, P 〈 0. 01 ） and 99. 3% （ Kappa = 0. 948 8, P 〈 0. 01 ） respectively. Drug resistance reports from these two systems showed similar results （Kappa = 0. 637 4, P 〈 0. 01 ）. The in-house detecting system identified 34 novel mutations besides the ViroSeqTM drug resistance mutation database （ViroSeqTM software v2.7 ）. Two mutations, V179F and K238T, had significant effect on HIV drug resistance. Conclusions The in-house genotyping system is an accurate, cost-effective method and has a high concordance with commercial ViroSeqTM genotyping system. Database from the in-house assay was superior to this of the ViroSeqTM assay.

关 键 词：HIV-1 基因型 抗药性 病毒 微生物敏感性试验 

分 类 号：R440[医药卫生—诊断学] R512.91[医药卫生—临床医学]