中国2007-2009年HIV-1耐药基因型检测质量评估结果

Evaluation of proficiency of HIV-1 drug resistance genotyping testing in three consecutive years in China

目的对2007-2009年统一发放至各省,针对艾滋病病毒Ⅰ型(HIV-1)蛋白酶和反转录酶基因的耐药突变基因型外部质控品的检测结果,进行分析和检测技术能力评价。方法对3年共5次由各省提交的,应用实验室自建方法(In house)、Viroseq(雅培公司,Abbott)和TruGene(西门子公司,Siemens)检测系统获得的耐药基因型数据进行综合分析。质控盘由6份血浆(HIV-1B及B/C重组毒株)组成,覆盖病毒的蛋白酶和反转录酶基因区的野生型和耐药突变型。每份序列与共享序列比较,使用扣分制对各实验室的结果进行评价。结果除3家核心实验室外,累计有19个省23家省市级疾病预防控制中心(CDC)或医院自愿参加,共提交了95份结果,其中3家实验室使用ViroSeq检测系统;提交了9份结果;1家使用TruGene检测系统;提交了1份结果;其余85份使用in-house检测系统。考核品总的序列阳性率(获得合格序列率)为97.7%,其中的低载量(1 000拷贝/mL)样本均得到阳性结果;根据累计提交的序列结果看,耐药位点判读的完全一致率为78.9%(75/95),序列编辑分析的一致率也有76.8%(73/95)。参加能力验证的省市级实验室由2007年的15家增加到2009年下半年的23家,合格率(001PT)由73.3%上升到(004PT)95.2%,3次以上优秀的实验室有13家。结论目前的检测方法均可成功扩增中国主要亚型流行毒株。所有检测结果在各实验室间的一致性均较高,因此基因型耐药检测技术是可靠的,但各实验室检测能力也存在差异。我国具备耐药基因型检测能力的实验室数量不断增加,检测水平逐年提高。

Objective To evaluate the ability to perform HIV 1 drug resistance genotypic testing at provincial/ city laboratories. Methods Five proficiency panels were distributed to provincial/city laboratories from 2007 to 2009. Every panel consisted of six plasma samples with wide or drug resistant HIV-1 viruses, which included sub- type B and CRF07_BC strains. HIV-1 drug resistance genotypic tests were performed by using in-house, Viroseq or Trugene method. Sequences containing whole protease gene and partial reverse transcriptase gene and corresponding mutation lists were submitted. Each laboratory was scored on basis of the number of disagreements with the consen- sus sequences. Results Twenty three laboratories from provincial and municipal CDCs or hospitals which voluntari- ly participated in testing submitted a total of 95 results, including 9 sets of results using Viroseq method from three laboratories and one using Trugene method from one other laboratory. The overall successful rate of sequencing was 97.7%. It was notable that all samples with a lower viral load （1000 copies/ml） were successfully sequenced. Seventy-five out of 95 （78.9%） results were consistent with consensus sequences on drug resistance related mutations. Seventy-three out of 95 （76.8%） were found to be consistent with consensus sequences on base calling and sequence editing. The number of participating laboratories increased from 15 to 23 between 2007 and 2009. The passing rate was raised from 73.3% （001PT） to 95.2% （004TP）. Thirteen labora- tories performed excellently for ≥3 times. Conclusion The present HIV1 drug resistance genotyping meth ods can be successfully used on strains prevailing in China. The results from participating laboratories were highly consistent. It is suggested that the present methods are reliable despite differences exiting among laborato- ries. The network of HIV drug resistance genotyping laboratories is expanding year by year.