A Mutation at Codon 516 in the rpoB Gene of Mycobacterium leprae Confers Resistance to Rifampin

Zhang, Liangfen; Namisato, Masako; Matsuoka, Masao

doi:10.1489/1544-581x(2004)72<468:amacit>2.0.co;2

Cited by 18 publications

(13 citation statements)

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“…Signals obtained by the LDS-DA with: a susceptible strain, Thai-53; strain Zensho-4, with mutation from ACC to ATC at codon 53 in the folP1 gene, from TCG to TTG at codon 425 in the rpoB gene and from GCA to GTA at codon 91 in the gyrA gene; strain Kusatsu-6, with mutation from CCC to CTC at codon 55 in the folP1 gene and from GAT to TAT at codon 410 in the rpoB gene; and strain Zensho-9, with mutation from CCC to CTC at codon 55 in the folP1 gene and from CAC to TAC at codon 420 in the rpoB gene. 53, Zensho-4 (Matsuoka et al, 2000) and two other strains with known nucleotide mutations (Zhang et al, 2004;Maeda et al, 2001). In Thai-53, which is susceptible to dapsone, rifampicin and ofloxacin, positive signals were observed on all of the wild-type spots.…”

Section: Resultsmentioning

confidence: 99%

“…A total of Simple detection of drug-resistant M. leprae 106 isolates without mutation in the rpoB gene and 63 isolates without mutation the gyrA gene were susceptible to rifampicin and ofloxacin, respectively. All isolates resistant to rifampicin or ofloxacin harboured mutations in the DRDR of rpoB or gyrA, respectively Williams et al, 1994Williams et al, , 2001Cambau et al, 1997Cambau et al, , 2001Matsuoka et al, 2000Matsuoka et al, , 2003Maeda et al, 2001;Zhang et al, 2004). Resistance of M. leprae to dapsone in the mouse footpad is classified into three degrees, namely, low, intermediate and high.…”

Section: Discussionmentioning

confidence: 99%

“…The molecular mechanism of rifampicin resistance was first demonstrated in Escherichia coli and thereafter in M. leprae . Rifampicin resistance is strongly correlated with mutations in the rpoB gene, encoding the b subunit of RNA polymerase Williams et al, 1994Williams et al, , 2001Matsuoka et al, 2000Matsuoka et al, , 2003Cambau et al, 2001;Maeda et al, 2001;Zhang et al, 2004). Resistance to fluoroquinolones has been proved to correlate with IP: 54.202.233.140…”

Section: Introductionmentioning

confidence: 99%

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A novel method for simple detection of mutations conferring drug resistance in Mycobacterium leprae, based on a DNA microarray, and its applicability in developing countries

Matsuoka

Aye

Kyaw

et al. 2008

Journal of Medical Microbiology

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A simple method to detect mutations in the genome of Mycobacterium leprae that confer resistance to key drugs for leprosy was exploited on the basis of a reverse hybridization system. A series of oligonucleotide probes corresponding to each mutation in the folP1, rpoB and gyrA genes for dapsone, rifampicin and ofloxacin resistance, respectively, were selected and fixed on a glass slide as capture probes, to develop a DNA microarray termed the leprosy drug susceptibility-DNA microarray (LDS-DA). Mutations in clinical isolates of M. leprae were successfully identified by the LDS-DA. Feasibility studies were conducted to evaluate the performance of the LDS-DA in two developing countries, Myanmar and the Philippines. The high concordance of the results obtained by this method with the results of nucleotide sequencing strongly supports the applicability of the LDS-DA as a drug susceptibility test in place of sequencing, a time-consuming and costly procedure. This is a rapid and simple method for the simultaneous susceptibility testing of three front-line drugs for leprosy, and solves the problems of previously reported methods.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A novel method for simple detection of mutations conferring drug resistance in Mycobacterium leprae, based on a DNA microarray, and its applicability in developing countries

Matsuoka

Aye

Kyaw

et al. 2008

Journal of Medical Microbiology

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“…These changes are determined by mutations in the drug resistance-determining region (DRDR) in the folP1, rpoβ, and gyrA genes which have demonstrated to confer resistance to dapsone, rifampicin, and ofloxacin, respectively. By defining resistance mechanisms it is possible to use DNAbased assays, namely PCR-direct DNA sequencing or reverse hybridization to examine susceptibility to these drugs (6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19).…”

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confidence: 99%

¿Es la resistencia de Mycobacterium leprae a los medicamentos un verdadero motivo de preocupación? Primera aproximación a la vigilancia molecular de pacientes colombianos multibacilares con tratamiento previo para lepra y sin él

et al. 2013

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Introduction:There is no information in Colombia on Mycobacterium leprae primary and secondary drug resistance in regards to the WHO-multidrug therapy regime. On the other hand, public health authorities around the world have issued various recommendations, one of which prompts for the immediate organization of resistance surveillance through simple molecular methods. Objective: To determine the prevalence of Mycobacterium leprae drug resistance to rifampicin, ofloxacin and dapsone in untreated and previously treated patients at the Centro Dermatológico Federico Lleras Acosta during the 1985-2004 period. Materials and methods: We conducted a retrospective study which included multibacillary patient biopsies through elective sampling: 381 of them from new patients and 560 from previously treated patients. Using a microtome, we obtained six slides from each skin biopsy preserved in paraffin, and we extracted M. leprae DNA. We amplified three molecular targets through PCR and obtained the patterns of drug resistance to dapsone, rifampicin and ofloxacin by reverse hybridization. Finally, we collected epidemiological, clinical and demographical data for analyses. Results: From 941 samples under study, 4.14% of them were resistant to one or more drugs, and 5.77 and 3.04% had resistant genotypes in new and previously treated patients, respectively. Total resistance for each drug was 0.43% for dapsone, 3.19% for rifampicin and 1.17% for ofloxacin. We found statistically significant differences for rifampicin and for the total population when comparing the results from untreated versus previously treated patients. Two thirds of the resistant samples were resistant to rifampicin alone or combined. Conclusions:The standard multidrug therapy schemes continue being effective for leprosy cases; however, it is necessary to guarantee adherence and regularity. Surveillance to drug resistance in new and previously treated leprosy cases should be established.

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“…Rifampicin (RIF) resistance is caused by mutations in the β-subunit of RNA polymerase, a target of RIF encoded by the rpoB gene (Zhang et al, 2004). More than 95% of the resistant strains harbor mutations in an 81-bp hot-spot region (codons 507 to 533) of the rpoB gene, named the RIFresistant determining region (RDR).…”

Section: Introductionmentioning

confidence: 99%

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

Wang¹,

Kim

et al. 2015

J Microbiol.

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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 -subunit of the RNA polymerase. The PCR-REBA simultaneously detects both 6 wild-type regions and 5 different mutations (507 AGC, 513 GTG, 516 TAT, 531 ATG, and 531 TTC) 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 531 ATG 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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A Mutation at Codon 516 in the rpoB Gene of Mycobacterium leprae Confers Resistance to Rifampin

Cited by 18 publications

References 9 publications

A novel method for simple detection of mutations conferring drug resistance in Mycobacterium leprae, based on a DNA microarray, and its applicability in developing countries

A novel method for simple detection of mutations conferring drug resistance in Mycobacterium leprae, based on a DNA microarray, and its applicability in developing countries

¿Es la resistencia de Mycobacterium leprae a los medicamentos un verdadero motivo de preocupación? Primera aproximación a la vigilancia molecular de pacientes colombianos multibacilares con tratamiento previo para lepra y sin él

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

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