Application of Genome Sequencing Analysis for Genotyping of Clinically Isolated Strains of Mycobacterium Leprae / 中华皮肤科杂志

ABSTRACT

Objective To genotype the clinically isolated strains of M.leprae by genome sequencing analysis. Methods PCR amplification was used to produce the 200 bp partial rpoT gene fragments from 2 standard strains and the isolated strains of M. leprae isolated from clinical specimens in 7 areas of China. The fragments were sequenced by BigDye Terminator Cycle Sequencing Reaction kit. Results① 12 rpoT－ 91/97bp DNA fragments and 2 rpoT－ 194/200 bp DNA fragments were obtained from 13 clinically isolated strains of M. leprae by PCR amplification;② based upon genome sequencing analysis, a component of 3－ copy or 4－ copy“ GACATC” repeat sequence was found in the nucleotide sequence of rpoT－ 194/200 bp DNA fragment. Conclusion The genome sequencing analysis can be used to objectively and accurately genotype M.leprae, and it is a useful tool for epidemiological study on transmission and infection of leprosy. However, the longer DNA fragments are necessary when sequencing analysis is conducted. Therefore this method needs further improvement because only the shorter DNA fragments amplified from paraffin－ embedded tissue.