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.