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

Sally M. McFall*, Robin L. Wagner, Sujit R. Jangam, Douglas H. Yamada, Diana Hardie, David M. Kelso

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)37-42
Number of pages6
JournalJournal of Virological Methods
Volume214
DOIs
StatePublished - Mar 5 2015

Keywords

  • Early infant diagnosis
  • HIV
  • Polymerase chain reaction
  • Provirus

ASJC Scopus subject areas

  • Virology

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