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Performance of PCR-reverse blot hybridization assay for detection of rifampicin-resistant Mycobacterium leprae

 Hyeyoung Wang  ;  Hyunjung Kim  ;  Yeun Kim  ;  Hyeeun Bang  ;  Jong Pill Kim  ;  Joo Hwan Hwang  ;  Sang Nae Cho  ;  Tae Ue Kim  ;  Hyeyoung Lee 
 Journal of Microbiology, Vol.53(10) : 686-693, 2015 
Journal Title
 Journal of Microbiology 
Issue Date
Amino Acid Sequence ; Antibiotics, Antitubercular/pharmacology* ; Base Sequence ; DNA-Directed RNA Polymerases/genetics ; Drug Resistance, Bacterial ; Humans ; Molecular Sequence Data ; Mutation ; Mycobacterium leprae/drug effects* ; Mycobacterium leprae/genetics* ; Mycobacterium leprae/isolation & purification ; Nucleic Acid Hybridization/methods* ; Polymerase Chain Reaction/methods* ; Rifampin/pharmacology* ; Sensitivity and Specificity
Mycobacterium leprae ; PCR-REBA (Reverse blot hybridization assay) ; molecular diagnosis ; rifampicin
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 -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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1. College of Medicine (의과대학) > Dept. of Microbiology (미생물학교실) > 1. Journal Papers
Yonsei Authors
Cho, Sang Nae(조상래)
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