A handheld HIV detection platform using paper-based sample preparation and real-time isothermal amplification.

Abstract

Early and accurate diagnosis of human immunodeficiency virus (HIV) infection is essential for timely initiation of antiretroviral therapy (ART) and prevention of new infections. However, conventional nucleic-acid-based tests for HIV detection require sophisticated laboratory equipment and trained personnel, which are often unavailable at the point-of-care (POC) or unaffordable in resource-limited settings. We report our development of a low-cost, integrated platform for POC testing of HIV. The platform integrates viral nucleic acid extraction on a paper substrate and reverse transcription loop-mediated isothermal amplification (RT-LAMP) in a portable, battery-powered heating device with real-time detection. The platform does not require laboratory infrastructure such as power outlets. The assay showed a detection limit of 30 copies/mL of HIV RNA in 140 μL human serum or 4 copies/reaction using 50 μL human serum, with no cross-reactivity with hepatitis C virus (HCV). We validated the platform using both plasma samples spiked with HIV and clinical samples from HIV-positive individuals, and compared it with standard laboratory assays based on polymerase chain reaction (PCR). These results demonstrate the feasibility of our platform for HIV testing at the POC.