Sequence‐specific detection method for reverse transcription, loop‐mediated isothermal amplification of HIV‐1

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‐seroconversion, 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. Published 2009 Wiley‐Liss, Inc.

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