HIV viral RNA extraction in wax immiscible filtration assisted by surface tension (IFAST) devices

Scott M. Berry, Alex J. Lavanway, Hannah M. Pezzi, David J. Guckenberger, Meghan A. Anderson, Jennifer M. Loeb, David J. Beebe

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

The monitoring of viral load is critical for proper management of antiretroviral therapy for HIV-positive patients. Unfortunately, in the developing world, significant economic and geographical barriers exist, limiting access to this test. The complexity of current viral load assays makes them expensive and their access limited to advanced facilities. We attempted to address these limitations by replacing conventional RNA extraction, one of the essential processes in viral load quantitation, with a simplified technique known as immiscible filtration assisted by surface tension (IFAST). Furthermore, these devices were produced via the embossing of wax, enabling local populations to produce and dispose of their own devices with minimal training or infrastructure, potentially reducing the total assay cost. In addition, IFAST can be used to reduce cold chain dependence during transportation. Viral RNA extracted from raw samples stored at 37°C for 1 week exhibited nearly complete degradation. However, IFAST-purified RNA could be stored at 37°C for 1 week without significant loss. These data suggest that RNA isolated at the point of care (eg, in a rural clinic) via IFAST could be shipped to a central laboratory for quantitative RT-PCR without a cold chain. Using this technology, we have demonstrated accurate and repeatable measurements of viral load on samples with as low as 50 copies per milliliter of sample.

Original languageEnglish
Pages (from-to)297-304
Number of pages8
JournalJournal of Molecular Diagnostics
Volume16
Issue number3
DOIs
StatePublished - May 2014

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Molecular Medicine

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