TY - JOUR
T1 - Smartphone-Imaged HIV-1 Reverse-Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) on a Chip from Whole Blood
AU - Damhorst, Gregory L.
AU - Duarte-Guevara, Carlos
AU - Chen, Weili
AU - Ghonge, Tanmay
AU - Cunningham, Brian T.
AU - Bashir, Rashid
N1 - Funding Information:
We would like to thank Dr. Bobby Reddy, Jr. for helpful discussions. We would also like to thank Dr. Bruce K. Brown and the NIH AIDS Reagent Program for their support and helpful discussions, though no program reagents were specifically used for this publication. Our work was supported by funding from the National Institutes of Health (NIH)Exploratory/Developmental Grant (R21) (AI106024). Gregory L. Damhorst is supported by a Ruth L. Kirschstein National Research Service Award for Individual Predoctoral MD/PhD and Other Dual Doctoral Degree Fellows (F30) (AI109825).
PY - 2015/9/1
Y1 - 2015/9/1
N2 - 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.
AB - 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.
KW - human immunodeficiency virus (HIV)
KW - loop-mediated isothermal amplification
KW - point-of-care
KW - smartphone
KW - viral load
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U2 - 10.15302/J-ENG-2015072
DO - 10.15302/J-ENG-2015072
M3 - Article
C2 - 26705482
AN - SCOPUS:85002622476
VL - 1
SP - 324
EP - 335
JO - Engineering
JF - Engineering
SN - 2095-8099
IS - 3
ER -