Development and Validation of the MAST ISOPLEX Kit for Simultaneous Detection of Shiga Toxin/Verotoxin 1 and 2 () with Inhibition Control (IC) in a Rapid Loop-Mediated Isothermal Amplification (LAMP) Multiplex Assay.

Loop-mediated isothermal amplification (LAMP) is a cost-effective, rapid, and highly specific method of replicating nucleic acids. Adding multiple targets into a single LAMP assay to create a multiplex format is highly desirable for clinical applications but has been challenging due to a need to develop specific detection techniques and strict primer design criteria. This study describes the evaluation of a rapid triplex LAMP assay, MAST ISOPLEX, for the simultaneous detection of Shiga toxin/verotoxin 1 and 2 ( and ) genes in verotoxigenic () isolates with inhibition control (IC) synthetic DNA using a single fluorophore-oligonucleotide probe, MAST ISOPLEX integrated into the primer set of each target.