HIV-1 DNA评价盘的构建及两种国产检测试剂性能评价

Performance evaluation of two kinds of HIV-1 DNA detection reagents

目的构建1型艾滋病病毒(HIV-1)脱氧核糖核酸(DNA)全血和干血斑评价盘,并对两种国产HIV-1 DNA检测试剂(试剂A和试剂B)性能进行实验室评价。方法从中国疾病预防控制中心性病艾滋病预防控制中心参比实验室样本库中筛选全血和干血斑样本,构建HIV-1 DNA全血和干血斑评价盘,评价盘包含阳性参考品20份,阴性参考品20份,最低检测限参考品5份,精密度参考品2份;并从云南省德宏州收集临床样本,通过评价盘和临床样本对两种试剂的敏感性、特异性、最低检测限和精密度进行评价。结果从云南省德宏州疾病预防控制中心和佑安医院共收集临床样本278份(全血和对应干血斑各278份),包含207份阳性样本和71份阴性样本。阳性样本中,106份未接受抗病毒治疗(ART),101份已接受ART。全血评价盘结果显示,两种试剂的阴、阳性样本符合率20/20,最低检测限均为4/5,A试剂精密度为1.2%(CV1)和1.4%(CV2),B试剂精密度为2.8%(CV1)和1.3%(CV2)。干血斑评价盘结果显示,两种试剂阴、阳性样本符合率均为20/20,最低检测限均为3/5,A试剂精密度为2.5%(CV1)和3.4%(CV2),B试剂精密度为3.1%(CV1)和2.8%(CV2)。177份未进行ART临床样本评价结果显示,对于全血样本,两种试剂特异性和敏感性均为100%(71/71,106/106);对于干血斑样本,两种试剂特异性均为100%(71/71),试剂A敏感性为98.1%(104/106),试剂B敏感性为99.1%(105/106)。101份已接受ART样本评价结果显示,对于全血样本,试剂A和试剂B敏感性均为100%(101/101);对于干血斑样本,试剂A敏感性为91.1%(92/101),试剂B敏感性为90.1%(91/101)。结论两种HIV DNA检测试剂均能符合全血和干血斑评价盘要求;在检测未进行ART全血和干血斑临床样本时均表现良好,且试剂A与B之间无显著差异,两种国产HIV-1 DNA检测试剂均能满足HIV-1核酸检测要求。

Objective To construct the reference panel for HIV DNA and evaluate the performance of two kinds of HIV DNA detection reagents. Methods Samples were screened from sample database of the national center for AIDS/STD control and prevention to construct the reference panel for HIV DNA(including whole blood panel and DBS panel), consisting of 20 positive references, 20 negative references, 5 detection limit references and 2 accurate references. Clinical samples were collected from Dehong prefecture center for disease control and prevention and Youan hospital. Results A total of 278 clinical samples were collected, including 207 positive samples and 71 negative samples. Performance of the two reagents was evaluated by using the reference panel and the clinical samples. Both of the two reagents met the requirements of the reference panel for HIV DNA. Detection results of the 177 pre-ART samples revealed that specificity and sensitivity of the two reagents were 100%(71/71) and 100%(106/106) for whole blood samples. For DBS samples, specificity of the two reagents was 100%(71/71) while sensitivity of reagent A was 98.1%(104/106) and that of reagent B was 99.1%(105/106). According to the detection results of the 101 samples receiving ART, sensitivity of the two reagents were 100%(101/101) for whole blood samples. Sensitivity of reagent A was 91.1%(92/101) while reagent B was 90.1%(91/101) for DBS samples. Conclusion Both of the two reagents meet the requirements of the reference panel for HIV DNA. The clinical samples of whole blood and DBS samples without ART show good performance, and there is no significant difference between reagent A and B, indicating that both of the two HIV-1 DNA reagents can meet the demands of HIV-1 nuclear detection.