LMV/DETTD/OBRR/CBER/FDA, Bethesda, Maryland.
Voluntary Intern, Northwest High School, Germantown, Maryland.
J Med Virol. 2016 Jun;88(6):1092-7. doi: 10.1002/jmv.24431. Epub 2015 Dec 16.
The genetic diversity of Human Immunodeficiency Virus type-1(HIV-1) has been shown to affect the performance of Nucleic Acid Testing (NAT) of Human Immunodeficiency Virus type-1. Although, majority NAT assays were designed to detect the conserved regions of HIV-1 mutations at the primer or probe binding regions may lead to false negatives. In this study, we evaluated the feasibility of detecting two genomic targets for enhanced sensitivity. A total of 180 tests using HIV-1 VQA RNA quantitation standard, 240 tests using EQAPOL HIV-1 viral diversity subtype panel, and 30 clinical plasma samples from Cameroon were evaluated. The analysis was based on probit and hit rate. The genomic targets LOD estimated by PROBIT for the gag target was 118 cps/ml (95%CI 64 cps/ml lower bound), Pol or POL/LTR was at 40 cps/ml (95%CI 17, 16 cps/ml), LTR 45 cps/ml (95%CI 20 cps/ml lower bound), and Gag/LTR at 67.8 cps/ml (95%CI 32 cps/ml lower bound). For HIV-1 subtypes the overall reactivity was 55-100% when tested at 100 and 1000 cps/ml and combination of genomic targets detection increased the reactivity to 100%. The plasma samples evaluation showed LTR or pol/LTR combination yielded higher sensitivity for patients with lower viral load (<40 cps/ml). We conclude that detection of two HIV-1 genomic targets improved sensitivity for detection of genetically diverse HIV-1 strains.
人类免疫缺陷病毒 1 型（HIV-1）的遗传多样性已被证明会影响 HIV-1 核酸检测（NAT）的性能。尽管大多数 NAT 检测试剂盒的设计目的是检测 HIV-1 突变的保守区域，但引物或探针结合区域的突变可能导致假阴性。在这项研究中，我们评估了检测两个基因组靶标以提高灵敏度的可行性。使用 HIV-1 VQA RNA 定量标准共评估了 180 次测试，使用 EQAPOL HIV-1 病毒多样性亚型面板进行了 240 次测试，还使用来自喀麦隆的 30 份临床血浆样本进行了评估。分析基于概率单位和命中率。PROBIT 估计 gag 靶标 LOD 的基因组靶标为 118 cps/ml（95%CI 下限 64 cps/ml），Pol 或 POL/LTR 为 40 cps/ml（95%CI 下限 17，16 cps/ml），LTR 为 45 cps/ml（95%CI 下限 20 cps/ml），Gag/LTR 为 67.8 cps/ml（95%CI 下限 32 cps/ml）。对于 HIV-1 亚型，当在 100 和 1000 cps/ml 时进行测试时，总体反应性为 55-100%，并且检测两个基因组靶标的组合可将反应性提高到 100%。对血浆样本的评估表明，LTR 或 pol/LTR 组合对病毒载量较低（<40 cps/ml）的患者具有更高的灵敏度。我们得出结论，检测两个 HIV-1 基因组靶标可提高对遗传多样性 HIV-1 株的检测灵敏度。
