title: A Sweet Spot for Molecular Diagnostics: Coupling Isothermal Amplification and Strand Exchange Circuits to Glucometers.
abstract: Strand exchange nucleic acid circuitry can be used to transduce isothermal nucleic acid amplification products into signals that can be readable on an off-the-shelf glucometer. Loop-mediated isothermal amplification (LAMP) is limited by the accumulation of non-specific products, but nucleic acid circuitry can be used to probe and distinguish specific amplicons. By combining this high temperature isothermal amplification method with a thermostable invertase, we can directly transduce Middle-East respiratory syndrome coronavirus and Zaire Ebolavirus templates into glucose signals, with a sensitivity as low as 20-100 copies/μl, equating to atto-molar (or low zepto-mole). Virus from cell lysates and synthetic templates could be readily amplified and detected even in sputum or saliva. An OR gate that coordinately triggered on viral amplicons further guaranteed fail-safe virus detection. The method describes has potential for accelerating point-of-care applications, in that biological samples could be applied to a transducer that would then directly interface with an off-the-shelf, approved medical device.
authors.affiliation: Center for Systems and Synthetic Biology, The University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Du
authors.affiliation: Applied Research Laboratories, The University of Texas at Austin, Austin, TX 78758, USA.
authors.forename: Randall A
authors.initials: RA
authors.lastname: Hughes
authors.affiliation: Center for Systems and Synthetic Biology, The University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Sanchita
authors.initials: S
authors.lastname: Bhadra
authors.affiliation: Department of Chemsitry, The University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Yu Sherry
authors.initials: YS
authors.lastname: Jiang
authors.affiliation: Center for Systems and Synthetic Biology, The University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Ellington
authors.affiliation: Center for Systems and Synthetic Biology, The University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Bingling
authors.initials: B
authors.lastname: Li
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D029043:Ebolavirus / Q000737:chemistry* / Q000235:genetics; D005947:Glucose / Q000032:analysis*; D021141:Nucleic Acid Amplification Techniques; D045473:Severe acute respiratory syndrome-related coronavirus / Q000737:chemistry* / Q000235:genetics; D043324:beta-Fructofuranosidase / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D043324:beta-Fructofuranosidase; D005947:Glucose