Journal of Microbiology

Research Support, Non-U.S. Gov't

Performance of PCR-reverse blot hybridization assay for detection of rifampicin-resistant Mycobacterium leprae: Hye-young Wang , Hyunjung Kim , Yeun Kim , Hyeeun Bang , Jong-Pill Kim , Joo Hwan Hwang , Sang-Nae Cho , Tae Ue Kim , Hyeyoung Lee; J. Microbiol. 2015;53(10):686-693. Published online October 2, 2015; DOI: https://doi.org/10.1007/s12275-015-5057-9

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Drug resistance in Mycobacterium leprae is a significant problem in countries where leprosy is endemic. A sensitive, specific, and high-throughput reverse blot hybridization assay (REBA) for the detection of genotypic resistance to rifampicin (RIF) was designed and evaluated. It has been shown that resistance to RIF in M. leprae involves mutations in the rpoB gene encoding the β-subunit of the RNA polymerase. The PCR-REBA simultaneously detects both 6 wild-type regions and 5 different mutations (507AGC, 513GTG, 516TAT, 531ATG, and 531TTC) including the most prevalent mutations at positions 507 and 531. Thirty-one clinical isolates provided by Korea Institute of Hansen’s Disease were analyzed by PCR-REBA with RIF resistance of rpoB gene. As a
result
, missense mutations at codons 507 AGC and 531ATG with 2-nucleotide substitutions were found in one sample, and a missense mutation at codon 516 TAT and ΔWT6 (deletion of 530-534) was found in another sample. These cases were confirmed by DNA sequence analysis. This rapid, simple, and highly sensitive assay provides a practical alternative to sequencing for genotypic evaluation of RIF resistance in M. leprae.

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Prediction of Y haplogroup by polymerase chain reaction-reverse blot hybridization assay
Sehee Oh, Jungho Kim, Sunyoung Park, Seoyong Kim, Kyungmyung Lee, Yang-Han Lee, Si-Keun Lim, Hyeyoung Lee
Genes & Genomics.2019; 41(3): 297. CrossRef

Detection of rifampin resistance mutation and its altered nucleotide sequences in mycobacterium leprae isolated from Korean patients with leprosy: Kim, Soon Ok , Kim, Min Joo , Chae, Gue Tae , Suh, Joo Won; J. Microbiol. 1996;34(3):236-240.

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Abstract: Rifampin is the most powerful drug for treating leprosy and tuberculosis today. It inhibits initiation and elonation of RNA transcription by binding to β-subunit of RNA polymerase, leading to kill mycobacteria. We isolated one variant strain of Mycobacterium leprae from 24 Korean leprosy patients who are less susceptible to rifampin or have suffered from relapse by polymerase chain reaction and single strand conformation polymorphism (PCR-SSCP) of the rpoB gene. Direct sequencing of the rpoB to Ser-464, Arg-465, Arg-467 and Ala-468. This is the first finding on rpoB gene mutation of M. leprae from Korean patients ; moreover the mutant type was found to be different from the previously reported cases in other countries.

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