Yufenyuy Ernest L, Parekh Bharat S
Division of Global HIV and TB, Center for Global Health, Centers for Disease Control and Prevention, Atlanta, Georgia.
AIDS Res Hum Retroviruses. 2018 Dec;34(12):1017-1027. doi: 10.1089/AID.2017.0279. Epub 2018 Sep 22.
Laboratory assays that can accurately distinguish recent (occurring within the past year) from long-standing (>1 year) HIV infection are crucial for understanding HIV transmission dynamics in a population. However, often these efforts are confounded by inaccurate HIV diagnosis and the presence of HIV-2 in the population being surveyed. This study describes development of a multiplex assay that can simultaneously perform HIV diagnosis, HIV serotyping, and detection of recent HIV-1 infection in a single well. HIV diagnosis and HIV-2 serotyping were accomplished by coupling beads with an HIV-1 p24-gp41 fusion protein and HIV-2 peptide from gp36 immunodominant region, respectively. HIV-1 recent infection detection was accomplished by coupling beads with limiting amounts of multi-subtype gp41 immunodominant protein, recombinant immunodominant region, group M (rIDR-M). Assay conditions, including concentration of coupled antigens, were systematically optimized using well-characterized specimens with known HIV-status (positive or negative), HIV-2 specimens, and recent or long-term HIV-1 classification based on LAg-Avidity enzyme immunoassay (EIA) in a stepwise manner. Beads were then combined in a multiplex assay to evaluate its performance using large panel of specimens (n = 1,500) that included HIV-1 positive (n = 570, recent = 78, long-term = 492), HIV-2 positive (n = 31), and seronegative individuals (n = 899). The diagnostic component of the assay performed with high sensitivity (99.8%) and specificity (99.7%), while the HIV-2 serotyping sensitivity and specificity were 96.7% and 100%, respectively. There was a high correlation (R = 0.84) between the LAg-Avidity EIA and the multiplex assay for recent infection detection. The assay showed high inter- and intra-assay reproducibility with %coefficient of variation of <10% in the dynamic range. The multiplex assay has the ability to diagnose HIV infection, perform serotyping, and detect and distinguish recent from long-term HIV infections, all in a single well. This novel assay has the potential to simplify HIV surveillance by reducing the multiple steps that are otherwise required.
能够准确区分近期（过去一年内发生）和长期（超过一年）HIV感染的实验室检测方法对于了解人群中的HIV传播动态至关重要。然而，这些工作常常因HIV诊断不准确以及被调查人群中存在HIV-2而受到干扰。本研究描述了一种多重检测方法的开发，该方法可以在单个孔中同时进行HIV诊断、HIV血清分型以及检测近期HIV-1感染。HIV诊断和HIV-2血清分型分别通过将珠子与HIV-1 p24-gp41融合蛋白以及来自gp36免疫显性区域的HIV-2肽偶联来完成。HIV-1近期感染检测通过将珠子与限量的多亚型gp41免疫显性蛋白、重组免疫显性区域M组（rIDR-M）偶联来完成。使用具有已知HIV状态（阳性或阴性）的特征明确的标本、HIV-2标本以及基于LAg-Avidity酶免疫测定（EIA）的近期或长期HIV-1分类，逐步系统地优化检测条件，包括偶联抗原的浓度。然后将珠子组合在多重检测中，使用大量标本（n = 1500）评估其性能，这些标本包括HIV-1阳性（n = 570，近期 = 78，长期 = 492）、HIV-2阳性（n = 31）和血清阴性个体（n = 899）。该检测方法的诊断部分具有高灵敏度（99.8%）和特异性（99.7%），而HIV-2血清分型的灵敏度和特异性分别为96.7%和100%。在近期感染检测方面，LAg-Avidity EIA与多重检测之间存在高度相关性（R = 0.84）。该检测方法在动态范围内显示出高的批间和批内重复性，变异系数百分比<10%。这种多重检测方法能够在单个孔中诊断HIV感染、进行血清分型以及检测和区分近期与长期HIV感染。这种新颖的检测方法有可能通过减少原本所需的多个步骤来简化HIV监测。
