title: Coupling Sensitive Nucleic Acid Amplification with Commercial Pregnancy Test Strips.
abstract: 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.
authors.affiliation: Institute for Cellular and Molecular Biology, Center for Systems and Synthetic Biology, Department of Chemistry, The University of Texas at Austin, Austin, TX, 78712, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Du
authors.affiliation: Institute for Cellular and Molecular Biology, Center for Systems and Synthetic Biology, Department of Chemistry, The University of Texas at Austin, Austin, TX, 78712, USA.
authors.forename: Arti
authors.initials: A
authors.lastname: Pothukuchy
authors.affiliation: Institute for Cellular and Molecular Biology, Center for Systems and Synthetic Biology, Department of Chemistry, The University of Texas at Austin, Austin, TX, 78712, USA.
authors.forename: Jimmy D
authors.initials: JD
authors.lastname: Gollihar
authors.affiliation: Institute for Cellular and Molecular Biology, Center for Systems and Synthetic Biology, Department of Chemistry, The University of Texas at Austin, Austin, TX, 78712, USA.
authors.forename: Armin
authors.initials: A
authors.lastname: Nourani
authors.affiliation: State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Jilin, P.R. China.
authors.forename: Bingling
authors.initials: B
authors.lastname: Li
authors.affiliation: Institute for Cellular and Molecular Biology, Center for Systems and Synthetic Biology, Department of Chemistry, The University of Texas at Austin, Austin, TX, 78712, USA.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Ellington
mesh_terms: D015415:Biomarkers / Q000032:analysis; D005260:Female; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D009696:Nucleic Acids / Q000032:analysis*; D019095:Point-of-Care Systems; D011247:Pregnancy; D011258:Pregnancy Tests
chemical_list: D015415:Biomarkers; D009696:Nucleic Acids
keywords: SNAP protein; human chorionic gonadotropin; loop-mediated isothermal amplification (LAMP); nucleic acid amplification; pregnancy test strips