Cross-subtype detection of HIV-1 using reverse transcription and recombinase polymerase amplification.
Cross-subtype detection of HIV-1 using reverse transcription and recombinase polymerase amplification.
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DOI：
10.1016/j.jviromet.2016.01.010
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发表时间：
2016-04
影响因子：
3.1
通讯作者：
Boyle DS
中科院分区：
文献类型：
--
作者：
Lillis L;Lehman DA;Siverson JB;Weis J;Cantera J;Parker M;Piepenburg O;Overbaugh J;Boyle DS
一种低复杂性的诊断测试，可以在护理点快速可靠地检测婴儿的艾滋病毒感染，可以促进早期治疗，改善结果。然而，许多婴儿艾滋病毒诊断只能在实验室环境中进行。酶聚合酶扩增（RPA）是一种等温扩增技术，可以在20分钟内快速扩增HIV-1多种亚型的前病毒DNA，而无需复杂的设备。在这项研究中，我们将逆转录（RT）添加到RPA中，以检测HIV-1 RNA和DNA。我们表明，这种RT-RPA HIV-1检测试剂盒的检测限分别为10至30个精确序列匹配的DNA或RNA拷贝。此外，在100个RNA或DNA拷贝时，该检测试剂盒检出了175个序列变体中的171个（97.7%），这些序列变体代表HIV-1 M组和O组的所有主要亚型和重组形式。这些数据表明，应用RT-RPA联合检测HIV-1病毒RNA和前病毒DNA可能被证明是一种快速准确诊断婴儿HIV的高灵敏度工具。
A low complexity diagnostic test that rapidly and reliably detects HIV infection in infants at the point of care could facilitate early treatment, improving outcomes. However, many infant HIV diagnostics can only be performed in laboratory settings. Recombinase polymerase amplification (RPA) is an isothermal amplification technology that can rapidly amplify proviral DNA from multiple subtypes of HIV-1 in under twenty minutes without complex equipment. In this study we added reverse transcription (RT) to RPA to allow detection of both HIV-1 RNA and DNA. We show that this RT-RPA HIV-1 assay has a limit of detection of 10 to 30 copies of an exact sequence matched DNA or RNA, respectively. In addition, at 100 copies of RNA or DNA, the assay detected 171 of 175 (97.7 %) sequence variants that represent all the major subtypes and recombinant forms of HIV-1 Groups M and O. This data suggests that the application of RT-RPA for the combined detection of HIV-1 viral RNA and proviral DNA may prove a highly sensitive tool for rapid and accurate diagnosis of infant HIV.