Smartphone-Imaged HIV-1 Reverse-Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) on a Chip from Whole Blood

Gregory L. Damhorst, Carlos Duarte-Guevara, Weili Chen, Tanmay Ghonge, Brian T. Cunningham, Rashid Bashir

Research output: Contribution to journalArticle

Abstract

Viral load measurements are an essential tool for the long-term clinical care of human immunodeficiency virus (HIV)-positive individuals. The gold standards in viral load instrumentation, however, are still too limited by their size, cost, and sophisticated operation for these measurements to be ubiquitous in remote settings with poor healthcare infrastructure, including parts of the world that are disproportionately affected by HIV infection. The challenge of developing a point-of-care platform capable of making viral load more accessible has been frequently approached but no solution has yet emerged that meets the practical requirements of low cost, portability, and ease-of-use. In this paper, we perform reverse-transcription loop-mediated isothermal amplification (RT-LAMP) on minimally processed HIV-spiked whole blood samples with a microfluidic and silicon microchip platform, and perform fluorescence measurements with a consumer smartphone. Our integrated assay shows amplification from as few as three viruses in a ~ 60 nL RT-LAMP droplet, corresponding to a whole blood concentration of 670 viruses per μL of whole blood. The technology contains greater power in a digital RT-LAMP approach that could be scaled up for the determination of viral load from a finger prick of blood in the clinical care of HIV-positive individuals. We demonstrate that all aspects of this viral load approach, from a drop of blood to imaging the RT-LAMP reaction, are compatible with lab-on-a-chip components and mobile instrumentation.

Original languageEnglish (US)
Pages (from-to)324-335
Number of pages12
JournalEngineering
Volume1
Issue number3
DOIs
StatePublished - Sep 1 2015

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Keywords

  • human immunodeficiency virus (HIV)
  • loop-mediated isothermal amplification
  • point-of-care
  • smartphone
  • viral load

ASJC Scopus subject areas

  • Computer Science(all)
  • Environmental Engineering
  • Chemical Engineering(all)
  • Materials Science (miscellaneous)
  • Energy Engineering and Power Technology
  • Engineering(all)

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