Department of Laboratory Medicine, All India Institute of Medical Sciences, Room No 11, 2nd Floor, Ansari Nagar, New Delhi, 110029, India.
Sci Rep. 2024 Apr 15;14(1):8700. doi: 10.1038/s41598-024-56164-5.
HIV infection has been a global public health threat and overall reported ~ 40 million deaths. Acquired immunodeficiency syndrome (AIDS) is attributed to the retroviruses (HIV-1/2), disseminated through various body fluids. The temporal progression of AIDS is in context to the rate of HIV-1 infection, which is twice as protracted in HIV-2 transmission. Q-PCR is the only available method that requires a well-developed lab infrastructure and trained personnel. Micro-PCR, a portable Q-PCR device, was developed by Bigtec Labs, Bangalore, India. It is simple, accurate, fast, and operationalised in remote places where diagnostic services are inaccessible in developing countries. This novel micro-PCR determines HIV-1 and HIV-2 viral load using a TruePrep™ extractor device for RNA isolation. Five ml blood samples were collected at the blood collection centre at AIIMS, New Delhi, India. Samples were screened for serology, and a comparison of HIV-1/2 RNA was done between qPCR and micro-PCR in the samples. The micro-PCR assay of HIV-RNA has compared well with those from real-time PCR (r = 0.99, i < 0.002). Micro-PCR has good inter and intra-assay reproducibility over a wide dynamic range (1.0 × 10-1.0 × 10 IU/ml). The linear dynamic range was 10-10 IU/ml. The clinical and analytical specificity of the assay was comparable, i.e., 100%. Intra-assay and inter-assay coefficients of variation ranged from 1.17% to 3.15% and from 0.02% to 0.46%, respectively. Moreover, due to the robust, simple, and empirical method, the Probit analysis has also been done for qPCR LODs to avoid uncertainties in target recoveries. The micro-PCR is reliable, accurate, and reproducible for early detection of HIV-1 and HIV-2 viral loads simultaneously. Thus, it can easily be used in the field and in remote places where quantification of both HIV-1/2 is not reachable.
HIV 感染一直是全球公共卫生威胁，总报告死亡人数约为 4000 万。获得性免疫缺陷综合征（AIDS）归因于逆转录病毒（HIV-1/2），通过各种体液传播。艾滋病的时间进展与 HIV-1 感染率有关，HIV-2 传播的感染率延长了一倍。实时荧光定量聚合酶链反应（Q-PCR）是唯一可用的方法，需要有完善的实验室基础设施和训练有素的人员。微-PCR 是一种由印度班加罗尔的 Bigtec Labs 开发的便携式 Q-PCR 设备。它简单、准确、快速，并在发展中国家无法获得诊断服务的偏远地区实施。这种新型微-PCR 使用 TruePrep™提取器设备分离 RNA 来确定 HIV-1 和 HIV-2 病毒载量。在印度新德里的 AIIMS 血液采集中心采集了 5ml 血液样本。对这些样本进行了血清学筛查，并在样本中比较了 qPCR 和微-PCR 对 HIV-1/2 RNA 的检测。HIV-RNA 的微-PCR 检测与实时 PCR（r = 0.99，i < 0.002）吻合良好。微-PCR 在宽动态范围内具有良好的内、间试验可重复性（1.0×10-1.0×10 IU/ml）。线性动态范围为 10-10 IU/ml。该检测方法的临床和分析特异性相当，即 100%。内、间试验变异系数分别为 1.17%至 3.15%和 0.02%至 0.46%。此外，由于该方法稳健、简单、经验性，还对 qPCR 的检测限进行了 Probit 分析，以避免目标回收率的不确定性。微-PCR 可用于早期同时检测 HIV-1 和 HIV-2 病毒载量，可靠、准确且可重复。因此，它可以很容易地在没有定量检测 HIV-1/2 的地区和野外使用。
