title: A point-of-care PCR test for HIV-1 detection in resource-limited settings.
abstract: 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.
authors.affiliation: Center for Innovation in Global Health Technologies, Biomedical Engineering, 2145 Sheridan Road E310, Northwestern University, Evanston, IL 60208-3107, USA. s-jangam@u.northwestern.edu
authors.forename: Sujit R
authors.initials: SR
authors.lastname: Jangam
authors.forename: Abhishek K
authors.initials: AK
authors.lastname: Agarwal
authors.forename: Kunal
authors.initials: K
authors.lastname: Sur
authors.forename: David M
authors.initials: DM
authors.lastname: Kelso
mesh_terms: D015374:Biosensing Techniques; D004279:DNA, Viral / Q000737:chemistry / Q000302:isolation & purification*; D015658:HIV Infections / Q000175:diagnosis / Q000821:virology; D015497:HIV-1 / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D007231:Infant, Newborn; D016133:Polymerase Chain Reaction
chemical_list: D004279:DNA, Viral