471939 Loop-Mediated Isothermal Amplification (LAMP): An Insight into Reaction Mechanism and Application in Point-of-Care Diagnostics
Although LAMP is increasingly used for diagnostic testing, current softwares for LAMP are limited to primer design, and unlike PCR, there is no known mechanistic description of the complete LAMP reaction. Here we present a kinetic model that explores all reaction intermediates and their evolution in respective reaction pathways. The kinetic parameters are estimated from literature and statistical mechanics calculation. The agent based model predicts around 15 side products along with the double loop DNA structure responsible for the autocatalytic amplification reaction. Some of these side products without the target sequence can lead to misrepresentation of the true amplification signal rendering LAMP as a semi-quantitative technique. The fundamental insights gained from our model are then used to determine optimal reaction parameters and efficient primer design. We leverage our new understandings to demonstrate multiplexed assay detection via the quencing of unincorporated amplification of signal reporters (QUASR) technique, enabling us to detect bright, unambiguous fluorescence signals with a simple smartphone based fluorimeter for point of care applications.
Sandia National Laboratories is a multi-program laboratory managed and operated by Sandia Corporation, a wholly owned subsidiary of Lockheed Martin Corporation, for the U.S. Department of Energy’s National Nuclear Security Administration under contract DE-AC04-94AL85000.