A multiplexed RT-LAMP assay for detection of group M HIV-1 in plasma or whole blood
A multiplexed RT-LAMP assay for detection of group M HIV-1 in plasma or whole blood
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DOI：
10.1016/j.jviromet.2018.02.012
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发表时间：
2018-05-01
影响因子：
3.1
通讯作者：
Owen, S. Michele
中科院分区：
文献类型：
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作者：
Curtis, Kelly A.;Morrison, Daphne;Owen, S. Michele
等温核酸扩增技术，例如逆转录环介导的等温扩增 (RT-LAMP)，具有适合开发可在 POC 中使用的快速、低成本 NAT 的特性。为了证明全球使用的实用性，需要进行研究来验证 RT-LAMP 在检测不同亚型方面的性能。在本研究中，我们设计并评估了多重 HIV-1 整合酶 RT-LAMP 引物，以检测 M 组内的亚型，以及 RNase P 阳性内部处理和扩增对照。使用一组代表主要循环亚型的 26 个病毒分离株，我们证明了亚型 A1、C、D、F1、F2、G、CRFO1_AE、CRF02_AG 和两种独特重组形式 (URF) 的所有分离株的检测。使用 M 组 HIV-1 整合酶和 RNase P 内部对照引物成功扩增了由每种亚型的一个代表性分离株创建的全血组。对从喀麦隆和乌干达获得的 61 份血浆样本进一步评估了 M 组 HIV-1 RT-LAMP 检测。序列保守的 M 组 HIV-1 RT-LAMP 引物与低成本扩增装置相结合，可以改善 POC 急性感染的诊断，并及时确认 HIV 状态。
Isothermal nucleic acid amplification techniques, such as reverse-transcription loop-mediated isothermal amplification (RT-LAMP), exhibit characteristics that are suitable for the development of a rapid, low-cost NAT that can be used at the POC. For demonstration of utility for global use, studies are needed to validate the performance of RT-LAMP for the detection of divergent subtypes. In this study, we designed and evaluated multiplexed HIV-1 integrase RT-LAMP primers to detect subtypes within group M, along with an RNase P positive internal processing and amplification control. Using a panel of 26 viral isolates representing the major circulating subtypes, we demonstrated detection of all isolates of subtypes Al, C, D, Fl, F2, G, CRFO1_AE, CRF02_AG, and two unique recombinant forms (URFs). A whole blood panel created with one representative isolate of each subtype was successfully amplified with the group M HIV-1 integrase and RNase P internal control primers. The group M HIV-1 RT-LAMP assay was further evaluated on 61 plasma specimens obtained from persons from Cameroon and Uganda. The sequence-conserved group M HIV-1 RT-LAMP primers, coupled to a low-cost amplification device, may improve diagnosis of acute infection at the POC and provide timely confirmation of HIV status.