01510nas a2200205   4500000000100000008004100001260001200042653001700054653001300071653001500084653000800099653000900107100001400116700001300130700001200143245010500155490000800260520102200268022001401290       2023                            d  c09/202310aEpidemiology10agenotype10atratamento10aPCR10aRFLP1 aDwivedi P1 aSharma M1 aSingh P00aMultiplex PCR-based RFLP assay for early identification of prevalent Mycobacterium leprae genotypes.0 v1073 a<p>Mycobacterium leprae is classified into four SNP genotypes and 16 subtypes (from 1A to 4P) that exhibit phylogeographical association reported from around the world. Among them, genotypes 1D and 3I represent more than 60% of M. leprae strains. Here, we report a new method for M. leprae genotyping which identifies the genotypes 1D and 3I by combining multiplex PCR amplification and restriction fragment length polymorphism (RFLP) of a M. leprae DNA amplicons using AgeI restriction enzyme. Agarose gel electrophoresis showed a deletion of 11 bp only among 3I genotypes by electrophoresis. When this multiplex PCR reaction is subjected to AgeI digestion, successful restriction digestion shows three bands for all the genotypes except 1D where only two bands were observed due to loss of restriction site. This method gives us the advantage of 1-step identification of the two most prevalent strains of M. leprae without using specialized equipments such as the Sanger sequencing system or quantitative PCR.</p>
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