Department of Clinical Virology, Christian Medical College, Vellore, Tamil Nadu, 632004, India.
Department of Infectious Diseases, Christian Medical College, Vellore, Tamil Nadu, 632004, India.
Curr HIV Res. 2022;20(4):274-286. doi: 10.2174/1570162X20666220609155237.
Currently, there is a global contemplation to end the AIDS epidemic by 2030. HIV-2 poses unique challenges to this end. The burden of HIV-2 is higher in resource-limited countries, and it is intrinsically resistant to NNRTI drugs. In addition, there is no FDA-approved plasma viral load assay to monitor disease progression and therapeutic efficacy. To overcome these challenges, we have developed and evaluated an in-house quantitative HIV-2 viral load assay.
Blood samples were collected from 28 HIV-2 treatment-naïve monoinfected individuals and tested using an in-house qPCR HIV-2 viral load assay. The extracted RNA was amplified using Quantifast pathogen + IC kit.
The in-house qPCR has a limit of detection of 695 copies/ml. The intra- and inter-assay variation (% CV) of the assay was 0.61 and 0.95, respectively. The in-house assay quantified HIV-2 NIBSC accurately (1000 IU) with a mean of 1952 copies/mL. Among the 28 samples tested by in-house qPCR assay, 11 (39.2%) samples were quantified, whereas 17 (60.7%) samples were not detected. In comparison with Altona RealStar HIV-2 RT PCR and Exavir Load RT assay, the results were 96.4% and 69.6% concordant, respectively. No significant (p = 0.99 and p = 0.13) difference in quantifying viral load between the three assays. Based on clinical and immunological (CD4) staging, the performance characteristics were comparable.
To the best of our knowledge, this is the first in-house qPCR developed in India. The performance characteristics of the in-house assay are comparable to the commercial assays, and they can be used assertively to monitor HIV-2 patients.
目前，全球正在思考如何在 2030 年终结艾滋病疫情。HIV-2 为此带来了独特的挑战。HIV-2 在资源有限的国家的负担更重，而且它对 NNRTI 类药物具有内在的耐药性。此外，目前还没有获得 FDA 批准的血浆病毒载量检测方法来监测疾病进展和治疗效果。为了克服这些挑战，我们开发并评估了一种内部的定量 HIV-2 病毒载量检测方法。
从 28 名未经治疗的 HIV-2 单感染个体中采集血液样本，并使用内部 qPCR HIV-2 病毒载量检测方法进行检测。提取的 RNA 使用 Quantifast 病原体+IC 试剂盒进行扩增。
内部 qPCR 的检测下限为 695 拷贝/ml。该检测方法的内和间检测变异系数（%CV）分别为 0.61 和 0.95。内部检测方法准确地定量了 HIV-2 NIBSC（1000IU），平均值为 1952 拷贝/ml。在使用内部 qPCR 检测方法检测的 28 个样本中，有 11 个（39.2%）样本被定量，而有 17 个（60.7%）样本未被检测到。与 Altona RealStar HIV-2 RT PCR 和 Exavir Load RT 检测方法相比，结果的符合率分别为 96.4%和 69.6%。三种检测方法在定量病毒载量方面无显著差异（p=0.99 和 p=0.13）。根据临床和免疫（CD4）分期，性能特征相当。
据我们所知，这是印度首次开发的内部 qPCR。内部检测方法的性能特征与商业检测方法相当，可以自信地用于监测 HIV-2 患者。
