Mathematical Modeling Of A Real-Time Isothermal Amplification Assay For Erwinia Amylovora

A general mathematical model that describes the temporal behavior of a real-time isothermal process used for nucleic acid amplification is derived. A monotonically-increasing fluorescence signal s(t) generated and measured during the amplification reaction can be modeled in the form of a logistic function of time that is completely described by three parameters (k, t50, and Smax), which may be readily estimated from experimentally acquired s(t) data. Experimental data obtained from a real-time loop-mediated isothermal amplification (LAMP) assay for the infectious pathogen Erwinia amylovora (E. amylovora) are used to illustrate and validate the mathematical model. Implementation of such a modeling approach can allow for the extraction of quantitative information from real-time LAMP data through parameter estimation techniques; this is demonstrated experimentally using real-time amplification data acquired using the real-time E. amylovora assay.