Premium
Coupling Sensitive Nucleic Acid Amplification with Commercial Pregnancy Test Strips
Author(s) -
Du Yan,
Pothukuchy Arti,
Gollihar Jimmy D.,
Nourani Armin,
Li Bingling,
Ellington Andrew D.
Publication year - 2017
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201609108
Subject(s) - loop mediated isothermal amplification , nucleic acid , biology , ebola virus , dna , rolling circle replication , computational biology , microbiology and biotechnology , biochemistry , genetics , polymerase , virus
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.