A paper and plastic device for performing recombinase polymerase amplification of HIV DNA.
Despite the importance of early diagnosis and treatment of HIV, only a small fraction of HIV-exposed infants in low- and middle-income countries are tested for the disease. The gold standard for early infant diagnosis, DNA PCR, requires resources that are unavailable in poor settings, and no point-of-care HIV DNA test is currently available. We have developed a device constructed of layers of paper, glass fiber, and plastic that is capable of performing isothermal, enzymatic amplification of HIV DNA. The device is inexpensive, small, light-weight, and easy to assemble. The device stores lyophilized enzymes, facilitates mixing of reaction components, and supports recombinase polymerase amplification in five steps of operation. Using commercially available lateral flow strips as a detection method, we demonstrate the ability of our device to amplify 10 copies of HIV DNA to detectable levels in 15 min. Our results suggest that our device, which is designed to be used after DNA extraction from dried-blood spots, may serve in conjunction with lateral flow strips as part of a point-of-care HIV DNA test to be used in low resource settings.
Department of Bioengineering, Rice University, Houston, TX, USA.
Brittany A
BA
Rohrman
Rebecca R
RR
Richards-Kortum
D004279:DNA, Viral / Q000032:analysis*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D059788:Dried Blood Spot Testing; D006678:HIV / Q000235:genetics*; D006801:Humans; D007223:Infant; D021141:Nucleic Acid Amplification Techniques; D010209:Paper; D010969:Plastics; D019095:Point-of-Care Systems; D045522:Recombinases / Q000378:metabolism*
D004279:DNA, Viral; D010969:Plastics; D045522:Recombinases; D004259:DNA-Directed DNA Polymerase