TY - JOUR
T1 - Coupling Sensitive Nucleic Acid Amplification with Commercial Pregnancy Test Strips
AU - Du, Yan
AU - Pothukuchy, Arti
AU - Gollihar, Jimmy D.
AU - Nourani, Armin
AU - Li, Bingling
AU - Ellington, Andrew D.
N1 - Funding Information:
This study was supported by the William and Melinda Gates Foundation (OPP1028808), the National Institutes of Health (U54EB015403), the National Security Science and Engineering Faculty Fellowship (FA9550-10-1-0169), the Welch Foundation (F-1654), and the Cancer Prevention Research Institute of Texas (RP140315).
Publisher Copyright:
© 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
PY - 2017/1/19
Y1 - 2017/1/19
N2 - 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-O6-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.
AB - 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-O6-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.
KW - SNAP protein
KW - human chorionic gonadotropin
KW - loop-mediated isothermal amplification (LAMP)
KW - nucleic acid amplification
KW - pregnancy test strips
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U2 - 10.1002/anie.201609108
DO - 10.1002/anie.201609108
M3 - Article
C2 - 27990727
AN - SCOPUS:85006970030
SN - 1433-7851
VL - 56
SP - 992
EP - 996
JO - Angewandte Chemie - International Edition
JF - Angewandte Chemie - International Edition
IS - 4
ER -