title: A simple and rapid DNA extraction method from whole blood for highly sensitive detection and quantitation of HIV-1 proviral DNA by real-time PCR.
abstract: Early diagnosis and access to treatment for infants with human immunodeficiency virus-1 (HIV-1) is critical to reduce infant mortality. The lack of simple point-of-care tests impedes the timely initiation of antiretroviral therapy. The development of FINA, filtration isolation of nucleic acids, a novel DNA extraction method that can be performed by clinic personnel in less than 2 min has been reported previously. In this report, significant improvements in the DNA extraction and amplification methods are detailed that allow sensitive quantitation of as little as 10 copies of HIV-1 proviral DNA and detection of 3 copies extracted from 100 μl of whole blood. An internal control to detect PCR inhibition was also incorporated. In a preliminary field evaluation of 61 South African infants, the FINA test demonstrated 100% sensitivity and specificity. The proviral copy number of the infant specimens was quantified, and it was established that 100 microliters of whole blood is required for sensitive diagnosis of infants.
authors: Center for Innovation in Global Health Technologies (CIGHT), Department of Biomedical Engineering, Northwestern University, Technical Institute E-310, 2145 Sheridan Rd., Evanston, IL 60208, USA. Electronic address: s-mcfall@northwestern.edu.
authors: Sally M
authors: SM
authors: McFall
authors: Center for Innovation in Global Health Technologies (CIGHT), Department of Biomedical Engineering, Northwestern University, Technical Institute E-310, 2145 Sheridan Rd., Evanston, IL 60208, USA; Pritzker School of Medicine, The University of Chicago, Chicago, IL 60637, USA.
authors: Robin L
authors: RL
authors: Wagner
authors: Center for Innovation in Global Health Technologies (CIGHT), Department of Biomedical Engineering, Northwestern University, Technical Institute E-310, 2145 Sheridan Rd., Evanston, IL 60208, USA.
authors: Sujit R
authors: SR
authors: Jangam
authors: Center for Innovation in Global Health Technologies (CIGHT), Department of Biomedical Engineering, Northwestern University, Technical Institute E-310, 2145 Sheridan Rd., Evanston, IL 60208, USA; Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA.
authors: Douglas H
authors: DH
authors: Yamada
authors: Virology Laboratory of the National Health Laboratory Services, Groote Schuur Hospital, and University of Cape Town, South Africa.
authors: Diana
authors: D
authors: Hardie
authors: Center for Innovation in Global Health Technologies (CIGHT), Department of Biomedical Engineering, Northwestern University, Technical Institute E-310, 2145 Sheridan Rd., Evanston, IL 60208, USA; Northwestern Global Health Foundation, Evanston, IL 60208, USA.
authors: David M
authors: DM
authors: Kelso
mesh_terms: D001769:Blood / Q000821:virology*; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods* / Q000592:standards; D013997:Time Factors; D019562:Viral Load / Q000379:methods
chemical_list: D004279:DNA, Viral
keywords: Early infant diagnosis; HIV; Polymerase chain reaction; Provirus