Molecular Detection of Mycobacterium Leprae Using RLEP Loop-Mediated Isothermal Amplification and a Restriction Enzyme to Ensure Amplification Specificity.

Abstract

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