Sequence-Specific Detection Method for Reverse Transcription, Loop-Mediated Isothermal Amplification of HIV-1
Sequence-Specific Detection Method for Reverse Transcription, Loop-Mediated Isothermal Amplification of HIV-1
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DOI：
10.1002/jmv.21490
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发表时间：
2009-06-01
影响因子：
12.7
通讯作者：
Owen, Michele
中科院分区：
文献类型：
--
作者：
Curtis, Kelly A.;Rudolph, Donna L.;Owen, Michele
由于时间和费用的限制，在护理点或资源有限的环境中进行艾滋病毒诊断带来了相当大的挑战。目前，基于核酸的检测是在感染后和血清转化前的窗口期诊断近期感染的唯一可靠方法，但这些检测仅适用于设备齐全的实验室环境。逆转录环介导等温扩增（RT-LAMP）技术具有理想的特性，可用于开发用于检测HIV DNA和RNA的快速、经济高效的核酸检测。本研究开发了一种序列特异性检测方法，可立即、肉眼可视化RT-LAMP产物，具有高灵敏度和特异性。将快速检测方法纳入HIV-1特异性RT-LAMP检测中，并使用来自HIV-1感染个体的微量全血进行验证。与最短的样品制备时间和一步等温扩增反应一起，序列特异性检测方法增加了RT-LAMP测定的整体通用性，并增强了在护理点或资源有限的地点使用的适用性。J. Med. Virol. 81：966-972，2009.出版2009 Wiley-Liss，Inc.匕首
HIV diagnosis at the point-of-care or in resource-limited settings poses considerable challenges due to time and cost limitations. Currently, nucleic acid-based tests are the only reliable method for diagnosing recent infections during the window period post-infection and pre-sero-conversion, but these tests are only suitable for well-equipped laboratory settings. The reverse transcription loop-mediated isothermal amplification (RT-LAMP) technology exhibits characteristics that are ideal for the development of a rapid, cost-effective nucleic acid-based test for detection of HIV DNA and RNA. In this study, a sequence-specific detection method was developed for immediate, naked-eye visualization of RT-LAMP products with high sensitivity and specificity. The rapid detection method was incorporated into the HIV-1-specific RT-LAMP assay and validated using minute volumes of whole blood from HIV-1-infected individuals. Together with the minimal sample preparation time and one-step, isothermal amplification reaction, the sequence-specific detection method adds to the overall versatility of the RT-LAMP assay and enhances the applicability for use at point-of-care or resource-limited sites. J. Med. Virol. 81:966-972, 2009. Publislied 2009 Wiley-Liss, Inc.dagger