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, Sangnae Cho, Tae Ue Kim, Hyeyoung Lee

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)686-693
Number of pages8
JournalJournal of Microbiology
Volume53
Issue number10
DOIs
Publication statusPublished - 2015 Oct 3

Fingerprint

Mycobacterium leprae
Rifampin
Polymerase Chain Reaction
Leprosy
Missense Mutation
Codon
Mutation
DNA-Directed RNA Polymerases
Korea
DNA Sequence Analysis
Drug Resistance
Genes
Nucleotides

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Applied Microbiology and Biotechnology

Cite this

Wang, Hye Young ; Kim, Hyunjung ; Kim, Yeun ; Bang, Hyeeun ; Kim, Jong Pill ; Hwang, Joo Hwan ; Cho, Sangnae ; Kim, Tae Ue ; Lee, Hyeyoung. / Performance of PCR-reverse blot hybridization assay for detection of rifampicin-resistant Mycobacterium leprae. In: Journal of Microbiology. 2015 ; Vol. 53, No. 10. pp. 686-693.
@article{4db5053a3bb345789809bc6c385d5cc1,
title = "Performance of PCR-reverse blot hybridization assay for detection of rifampicin-resistant Mycobacterium leprae",
abstract = "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.",
author = "Wang, {Hye Young} and Hyunjung Kim and Yeun Kim and Hyeeun Bang and Kim, {Jong Pill} and Hwang, {Joo Hwan} and Sangnae Cho and Kim, {Tae Ue} and Hyeyoung Lee",
year = "2015",
month = "10",
day = "3",
doi = "10.1007/s12275-015-5057-9",
language = "English",
volume = "53",
pages = "686--693",
journal = "Journal of Microbiology",
issn = "1225-8873",
publisher = "Microbiological Society of Korea",
number = "10",

}

Performance of PCR-reverse blot hybridization assay for detection of rifampicin-resistant Mycobacterium leprae. / Wang, Hye Young; Kim, Hyunjung; Kim, Yeun; Bang, Hyeeun; Kim, Jong Pill; Hwang, Joo Hwan; Cho, Sangnae; Kim, Tae Ue; Lee, Hyeyoung.

In: Journal of Microbiology, Vol. 53, No. 10, 03.10.2015, p. 686-693.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Performance of PCR-reverse blot hybridization assay for detection of rifampicin-resistant Mycobacterium leprae

AU - Wang, Hye Young

AU - Kim, Hyunjung

AU - Kim, Yeun

AU - Bang, Hyeeun

AU - Kim, Jong Pill

AU - Hwang, Joo Hwan

AU - Cho, Sangnae

AU - Kim, Tae Ue

AU - Lee, Hyeyoung

PY - 2015/10/3

Y1 - 2015/10/3

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84942844423&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84942844423&partnerID=8YFLogxK

U2 - 10.1007/s12275-015-5057-9

DO - 10.1007/s12275-015-5057-9

M3 - Article

VL - 53

SP - 686

EP - 693

JO - Journal of Microbiology

JF - Journal of Microbiology

SN - 1225-8873

IS - 10

ER -