Key Laboratory of Medical Virology, Ministry of Health, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China; College of Light Industry and Food Sciences, South China University of Technology, Guangzhou, China.
Key Laboratory of Medical Virology, Ministry of Health, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
Gene. 2014 May 15;541(2):123-8. doi: 10.1016/j.gene.2014.03.015. Epub 2014 Mar 12.
Accurate and rapid quantitation of Human immunodeficiency virus type 1 (HIV-1) RNA levels is a critical aspect in estimating the effect of antiviral therapy and establishing therapeutic schedule. Thus, for the first time, a rapid quantitative reverse transcription-loop-mediated isothermal amplification (RT-LAMP) was designed to quantitate HIV-1 RNA. The results showed that the dynamic range was from 2.5×10(2) to 10(7) copies with a coefficient of determination (R(2)) of 0.991, and the limit of detection of RT-LAMP by Probit analysis at the 95% detection level was 196 copies. The intra-assay coefficient of variation (CV) ranged from 0.67% to 2.08% at 10(7) copies and 7.25% to 12.97% at 250 copies. The CVs of inter-assay were 2.39% and 13.93% for the high and low copy numbers, respectively. No cross-reaction with Human immunodeficiency virus type 2 (HIV-2), Human T lymphotrophic virus type 1 (HTLV-1) and Hepatitis C virus (HCV) was observed and a good agreement between the RT-LAMP method and the real-time reverse transcription-polymerase chain reaction (qRT-PCR) test was achieved. This proposed RT-LAMP method could be useful for rapid diagnosis of high risk group and pharmacodynamic assessment of anti-HIV drug, especially in less-equipped laboratories of impoverished areas.
准确、快速地定量检测人类免疫缺陷病毒 1 型（HIV-1）RNA 水平是评估抗病毒治疗效果和制定治疗方案的关键。因此，我们首次设计了一种快速定量逆转录环介导等温扩增（RT-LAMP）方法来定量检测 HIV-1 RNA。结果表明，该方法的动态检测范围为 2.5×10(2)到 10(7)拷贝，决定系数（R(2)）为 0.991，95%检测水平下 Probit 分析的 RT-LAMP 检测限为 196 拷贝。在 10(7)拷贝和 250 拷贝时，批内变异系数（CV）范围分别为 0.67%至 2.08%和 7.25%至 12.97%。高、低拷贝数的批间 CV 分别为 2.39%和 13.93%。该方法与人类免疫缺陷病毒 2 型（HIV-2）、人类 T 淋巴细胞嗜病毒 1 型（HTLV-1）和丙型肝炎病毒（HCV）无交叉反应，与实时逆转录-聚合酶链反应（qRT-PCR）检测方法具有良好的一致性。该 RT-LAMP 方法可用于高危人群的快速诊断和抗 HIV 药物的药效评估，特别是在贫困地区设备简陋的实验室。
