Development of a LAMP assay using hybridization-based TaqMan-style probes (HyTaq) for simultaneous detection of wild-type and macrolide-resistant <i>Mycoplasma pneumoniae</i>

Macrolide-resistant <i>Mycoplasma pneumoniae</i> is a significant cause of treatment failure in community-acquired respiratory infections, particularly in children and adolescents. Early and accurate detection of resistance-associated mutations, such as A2063G and A2064G, is essential for guiding effective antibiotic therapy. In this study, we developed an MP-R HyTaq-based loop-mediated isothermal amplification assay capable of simultaneously detecting and discriminating wild-type strains and macrolide resistance mutations (A2063G and A2064G) in a single-tube, isothermal reaction. Its high specificity, rapid turnaround time, and minimal equipment requirements make this assay suitable for point-of-care testing in diverse clinical settings.