Rapid detection of HIV-1 by reverse-transcription, loop-mediated isothermal amplification (RT-LAMP).

ABSTRACT

A rapid, cost-effective diagnostic or confirmatory test for the detection of early HIV-1 infection is highly desired, especially for use in resource-poor or point-of-care settings. The reverse-transcription loop-mediated isothermal amplification (RT-LAMP) technology has been evaluated for the detection of HIV-1 DNA and RNA, using six RT-LAMP primers designed against highly conserved sequences located within the protease and p24 gene regions. Amplification from lab-adapted HIV-1 DNA and RNA was detected as early as 30 min, with maximum sensitivity of 10 and 100 copies per reaction, respectively, reached at 60 min. Comparable sensitivity was observed with extracted nucleic acid from plasma and blood samples of HIV-1-infected individuals. Furthermore, the RT-LAMP procedure was modified for the direct detection of HIV-1 nucleic acid in plasma and blood samples, eliminating the need for an additional nucleic acid extraction step and reducing the overall procedure time to approximately 90 min.