Molecular detection of Mycobacterium leprae using RLEP-LAMP and restriction enzyme to ensure amplification specificity.

Abstract

Early and accurate diagnosis of leprosy is important but remains a significant challenge till date. Loop-mediated isothermal amplification (LAMP) is an isothermal process for amplification of nucleic acids at constant temperature and has been used to develop field-friendly tests for many diseases. In the present study, we have described the development of a colorimetric LAMP assay targeting Mycobacterium leprae-specific 450 bp conserved region of the repeat sequences known as RLEP. Furthermore, the amplicons of LAMP were subjected to restriction analysis by the enzyme EcoRV for specificity. This method has the potential to become an accurate and efficient alternative to Sanger sequencing which is currently in use to validate the RLEP amplified products.