A point-of-care PCR test for HIV-1 detection in resource-limitedsettings

A low-cost, fully integrated sample-to-answer, quantitative PCR (qPCR) system that can be used for detection of HIV-1 proviral DNA in infants at the point-of-care in resource-limited settings has been developed and tested. The system is based on a novel DNA extraction method, which uses a glass fiber membrane, a disposable assay card that includes on-board reagent storage, provisions for thermal cycling and fluorescence detection, and a battery-operated portable analyzer. The system is capable of automated PCR mix assembly using a novel reagent delivery system and performing qPCR. HIV-1 and internal control targets are detected using two spectrally separated fluorophores, FAM and Quasar 670. In this report, a proof-of-concept of the platform is demonstrated. Initial results with whole blood demonstrate that the test is capable of detecting HIV-1 in blood samples containing greater than 5000 copies of HIV-1. In resource-limited settings, a point-of-care HIV-1 qPCR test would greatly increase the number of test results that reach the infants caregivers, allowing them to pursue anti-retroviral therapy.

► A point-of-care platform for PCR based HIV-1 testing is presented. ► Platform integrates DNA extraction, amplification and detection. ► A microfluidic assay card is used to assemble the reaction, thermocycle and detect fluorescence. ► A battery operated portable analyzer enables PCR based HIV-1 testing to be deployed in rural clinics. ► Point-of-care testing has the potential to reduce infant mortality by providing results to the caregivers in the samevisit.