Methylation-sensitive LAMP (MS-LAMP): assay design, readouts, and clinical translation in microbiology

Loop-mediated isothermal amplification (LAMP) is widely used when rapid nucleic acid detection with minimal instrumentation is required. This review focuses on methylation-sensitive LAMP (MS-LAMP), a narrower and still underdeveloped extension in which methylation status is converted into an amplification outcome. We concentrate on microbiology-relevant applications and discuss the two principal gating routes currently used: bisulfite conversion followed by methylation-discriminating primer design, and methylation-sensitive restriction enzyme digestion before amplification. Rather than reviewing LAMP broadly, we examine how methylation gating reshapes assay design, readout selection, upstream quality control, and result interpretation. We also distinguish direct MS-LAMP evidence from the larger supporting literature on microbial methylomes and general LAMP engineering. Current evidence suggests that the main translational bottlenecks lie upstream of amplification, particularly in template damage during bisulfite treatment, digestion specificity, control design, and performance in complex clinical matrices. We conclude that MS-LAMP remains promising but methodologically immature for routine microbiology, and that progress will depend on robust locus selection, process controls, predefined thresholds, and validation across clinically relevant specimen types.