Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 56, Issue 4, Pages 992-996Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201609108
Keywords
human chorionic gonadotropin; loop-mediated isothermal amplification (LAMP); nucleic acid amplification; pregnancy test strips; SNAP protein
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Funding
- William and Melinda Gates Foundation [OPP1028808]
- National Institutes of Health [U54EB015403]
- National Security Science and Engineering Faculty Fellowship [FA9550-10-1-0169]
- Welch Foundation [F-1654]
- Cancer Prevention Research Institute of Texas [RP140315]
- Bill & Melinda Gates Foundation - Grand Challenges Explorations Initiative [OPP1128792] Funding Source: researchfish
- Bill and Melinda Gates Foundation [OPP1128792] Funding Source: Bill and Melinda Gates Foundation
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The detection of nucleic acid biomarkers for point-of-care (POC) diagnostics is currently limited by technical complexity, cost, and time constraints. To overcome these shortcomings, we have combined loop-mediated isothermal amplification (LAMP), programmable toehold-mediated strand-exchange signal transduction, and standard pregnancy test strips. The incorporation of an engineered hCG-SNAP fusion reporter protein (human chorionic gonadotropin-O-6-alkylguanine-DNA alkyltransferase) led to LAMP-to-hCG signal transduction on low-cost, commercially available pregnancy test strips. Our assay reliably detected as few as 20 copies of Ebola virus templates in both human serum and saliva and could be adapted to distinguish a common melanoma-associated SNP allele (BRAF V600E) from the wild-type sequence. The methods described are completely generalizable to many nucleic acid biomarkers, and could be adapted to provide POC diagnostics for a range of pathogens.
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