Odari Eddy Okoth, Maiyo Alex, Lwembe Raphael, Gurtler Lutz, Eberle Josef, Nitschko Hans
Max von Pettenkofer-Institute for Hygiene and Medical Microbiology (MVPI), Ludwig-Maximilians-University, Pettenkofer Strasse 9a, 80336 Munich, Germany; Center for International Health (CIH) at the Ludwig-Maximilians-University, Leopoldstraße 7, 80802 Munich, Germany; Department of Medical Microbiology, Jomo Kenyatta University of Agriculture and Technology, P.O. Box 62 000, 00200 Nairobi, Kenya.
Kenya Medical Research Institute (KEMRI), Mbagathi, P.O. Box 54840, Nairobi, Kenya.
J Virol Methods. 2015 Feb;212:30-8. doi: 10.1016/j.jviromet.2014.10.012. Epub 2014 Nov 6.
The past decade has witnessed a dramatic increase of anti-retroviral treatment of human immunodeficiency virus (HIV) infected patients in many African countries. Due to costs and lack of currently available commercial viral load assays, insufficient attention has been paid to therapy monitoring through measurement of plasma viral load. This challenge of patient monitoring by tests as viral load, CD4 cell count, and finally HIV drug resistance could reverse achievements already made against HIV/AIDS infection. Loop-mediated isothermal amplification (LAMP) has been shown to be simple, rapid and cost-effective, characteristics which make this assay suitable for viral load monitoring in resource limited settings. This paper describes a revised LAMP assay using primers in the HIV-1 integrase region. The assay can be used for semi-quantitative measurement of HIV-1 group M viral load. The lower limit of detection (LLOD) was determined as 1200copies/mL and lower limit of quantitation (LLOQ) at 9800copies/mL. Sensitivities of 82 and 86% (in 135 and 99 plasma samples respectively from Kenya) and 93% (in 112 plasma samples from Germany) and specificities of 99 and 100% were realized. HIV-1 group O and HIV-2 virus samples were not detected. This LAMP assay has the potential for semi-quantitation of HIV-1 group M viral load in resource limited countries. There is still a need for further improvement by refinement of primers in respect to detection of HIV-1 group M non-B virus.
在过去十年中，许多非洲国家接受抗逆转录病毒治疗的人类免疫缺陷病毒（HIV）感染患者数量急剧增加。由于成本以及目前缺乏可用的商业病毒载量检测方法，通过检测血浆病毒载量进行治疗监测受到的关注不足。通过病毒载量、CD4细胞计数以及最终的HIV耐药性检测来监测患者的这一挑战，可能会使在抗击HIV/AIDS感染方面已经取得的成果付诸东流。环介导等温扩增技术（LAMP）已被证明具有操作简单、快速且成本效益高的特点，这些特性使得该检测方法适用于资源有限环境下的病毒载量监测。本文描述了一种使用HIV-1整合酶区域引物的改良LAMP检测方法。该检测方法可用于HIV-1 M组病毒载量的半定量检测。检测下限（LLOD）确定为1200拷贝/毫升，定量下限（LLOQ）为9800拷贝/毫升。分别在来自肯尼亚的135份和99份血浆样本中实现了82%和86%的灵敏度，在来自德国的112份血浆样本中实现了93%的灵敏度，特异性分别为99%和100%。未检测到HIV-1 O组和HIV-2病毒样本。这种LAMP检测方法有潜力在资源有限的国家对HIV-1 M组病毒载量进行半定量检测。在检测HIV-1 M组非B型病毒方面，仍需要通过优化引物来进一步改进。
