Mycobacterium tuberculosis mtb drug resistance is a global concern. Dec 21, 2015 rapid antibioticresistance predictions from genome sequence data for staphylococcus aureus and mycobacterium tuberculosis. Moreover, multiple drug resistant mdr, extensively drug resistant xdr, and totally drug resistant tdr mtb cases are on the rise in developing countries like india. Detection of a novel mutation in the rpob gene in a multidrug. Based on previous work, we hypothesized that changes in the expression of cell wall molecules in mtb mutant rpob 526d lead to changes in cell wall permeability and to. Genetic evaluation of relationship between mutations in rpob. Genetic evaluation of relationship between mutations in rpob and. Molecular evidence of lateral gene transfer in rpob gene of mycobacterium yongonense strains via. Resistance profiles and rpob gene mutations of mycobacterium tuberculosis isolates in taiwan. Get a printable copy pdf file of the complete article 1.
Lipf increases rifampicin and streptomycin sensitivity in. Introduction of xpert mtbrif assay has revolutionalised the diagnosis of tuberculosis tb by simultaneously detecting the bacteria and resistance to rifampicin rif, a surrogate marker for multidrug resistant tb mdrtb as well as one of the principal firstline antituberculosis drugs. Rifampicin resistance and mutation of the rpob gene in. He had a long history of chest infections and diabetes mellitus. With the widespread use of rifampicin and isoniazid, bacterial resistance has become a growing problem. Mutations in the rpob gene of rifampicinresistant mycobacterium. Using 39 clinical isolates of mycobacterium tuberculosis strains with a broad range of susceptibility to rifampicin. Mutations in the rpob gene of multidrugresistant mycobacterium. Characterization of mutations conferring resistance to rifampin in. A portion of the mycobacterium tuberculosis gene encoding the beta subunit of rna polymerase rpob was amplified by pcr using degenerate oligonucleotides and used as a hybridization probe to isolate plasmid clones carrying the entire rpob gene of m. The genome features file from ncbi was used to create a bed file to determine the geneorf startstop positions.
Resistance levels and rpob gene mutations among in vitroselected rifampinresistant mycobacterium tuberculosis mutants. Relationship between antimycobacterial activities of. Investigation of cases with discrepant susceptibility results pdf paperity. Mycobacterium bovis and the bacillus calmetteguerin strain. Rapid genotypic detection of rpob and katg gene mutations in. We also evaluated the correlation between the level of resistance to rifampicin, rifabutin and krm1648 and genetic alterations in the rpob gene. Jan 16, 20 the present study was undertaken to investigate the type and frequency of rpob and katg mutations in rifampicin rif and isoniazid inh resistant strains respectively of mycobacterium tuberculosis mtb circulating in northern india and to explore the utility of multiplex. Mutation analysis of mycobacterial rpob genes and rifampin.
Dec 01, 2006 the aim of this study was to investigate the frequency, location and type of rpob mutations in mycobacterium tuberculosis isolated from patients in belarus. Tuberculosis cases are increasing every year in belarus. Pdf characterization of rpob mutations in rifampin. A method for the molecular determination of rifampin resistance in these two mycobacteria would be clinically valuable, but the relationship between the mutations and susceptibility to rifampin must be clarified before its use. Dst centre of excellence for biomedical tuberculosis research. Rifampicin resistance is mediated solely by mutations in the rifampicinresistance determining region of the rpob gene, coding for the essential. Mutations in the rpob gene of rifampinresistant mycobacterium tuberculosis isolates from eastern poland. Determination of the minimal acidinducible promoter region of the lipf gene from mycobacterium tuberculosis. Detection of a novel mutation in the rpob gene in a. The objective of this study was to investigate the crossresistance between rifampin rif and rifabutin rfb among clinical mycobacterium tuberculosis mtb isolates, and the correlations between specific rpob mutations and the minimum inhibitory concentrations mics of rif and rfb. Mycobacterium tuberculosis rpob gene dna sequencing. Clinically relevant mutations in mycobacterial lepa cause. Restricted structural gene polymorphism in the mycobacterium tuberculosis complex indicates evolutionarily recent global dissemination. Free fulltext pdf articles from hundreds of disciplines, all in one place correlation between genotypic and phenotypic testing for resistance to rifampin in mycobacterium tuberculosis clinical isolates in haiti.
A rapid and sensitive chipbased assay for detection of rpob. Molecular characterization of multidrugresistant mycobacterium. However, compared with the slow traditional methods of. Rapid detection of rpob gene mutations conferring rifampin. The global incidence of drugresistant tuberculosis tb, particularly. Molecular evidence of lateral gene transfer in rpob gene. To determine the sensitivity of this method, we analyzed 35 rifr strains and 11 rifampinsusceptible rifs strains, using the dna sequencing of the core region of rpob for comparison. Rpob gene of mycobacterium tuberculosis pdf tuberculosis rpob gene was cloned into the shuttle plasmid pmv261 and electroporated into the lr223 strain of mycobacterium smegmatis, which is. Characterization of rpob mutations in rifampinresistant clinical mycobacterium tuberculosis isolates from greece article pdf available in journal of clinical microbiology 361.
Consensus numbering system for the rifampicin resistance. Recently, a novel species, mycobacterium yongonense dsm 45126t, was introduced and while it is phylogenetically related to mycobacterium intracellulare, it has a distinct rna polymerase. The rpob gene sequence showed that resistance was associated with rare mutations in each isol. Pcr procedures for rpob gene amplification aliquots of purified m. Frontiers altered mycobacterium tuberculosis cell wall. Magnitude of gene mutations conferring drug resistance in. Molecular evidence of lateral gene transfer in rpob gene of. An inhibitor of transcription in bacteria, tagetitoxin, also inhibits pep, showing that the complex found in plants is very similar to the homologous enzyme in bacteria.
The use of polymerase chain reactionsinglestranded conformational polymorphism pcrsscp to study rpob gene mutations in rifampinresistant rifr mycobacterium tuberculosis has yielded contradictory results. The aim of this study was to analyze mutations occurring in the rpob gene of mycobacterium tuberculosis mtb isolates from clinical samples of extrapulmonary tuberculosis eptb. However, the net effects of rpob mutations on mtb physiology, metabolism and function are not completely understood. The aim of the present study was to investigate the mutational characteristics of the drug. Mutations in rpob and katg genes in mycobacterium isolates. Rifabutin resistance associated with double mutations in rpob. Pdf molecular characterization of the rpob gene mutations. Exploring malditof mass spec for mycobacteria cap today. A total of 256 rifresistant isolates were included from the national tuberculosis clinical laboratory in china.
The situation of tb is made even worse by the rising emergence of drug resistant strains of mycobacterium tuberculosis. Additionally, the lack of relevant baseline information for the frequency of drugresistant tuberculosis tb gene mutations is a critical issue, and the incidence of this infection in the city of changchun has not investigated to date. Correlation between genotypic and phenotypic testing for. The rifampin resistance of mycobacterium lepraeand mycobacterium tuberculosisresults from a mutation in the rpobgene, encoding the. Links huitric e, werngren j, jureen p, hoffner s 2006. Pdf there is a geographic distribution of mycobacterium tuberculosis strains with various rpob gene mutations that account for rifampin resistance. Resistance of mycobacterium tuberculosis to rifampin rmp, mediated by mutations in the rpob gene coding for the betasubunit of rna. These two primers were prepared to completely match the m.
Molecular characterization of the rpob gene mutations of mycobacterium tuberculosis isolated from china article pdf available in journal of tuberculosis research 0101. Resistance profiles and rpob gene mutations of mycobacterium. A total of 256 rifresistant isolates were included from the national tuberculosis clinical laboratory in. The former has not yet been cultured on artificial media. Characterization of rpob mutations in rifampicinresistant mycobacterium tuberculosis isolated in china. Multiple rpob mutants of mycobacterium tuberculosis and. Detection of mutations in rpob gene of clinically isolated m. Characterization of rpob mutations associated with rifampin resistance in mycobacterium tuberculosis from eastern china. The mycobacterium tuberculosis complex mtc or mtbc is a genetically related group of mycobacterium species that can cause tuberculosis in humans or other animals. Molecular characterization of the rpob gene mutations of mycobacterium tuberculosis isolated from china. The bst eii fragment 1716 bp of rpob gene can be easily amplified based on genomic dna isolated from investigated m.
Deciphering the biology of mycobacterium tuberculosis from. Recently, a novel species, mycobacterium yongonense dsm 45126 t, was introduced and while it is phylogenetically related to mycobacterium intracellulare, it has a distinct rna polymerase. Mycobacterium tuberculosis mtb rpob mutations are associated with global metabolic remodeling. Frontiers rifabutin resistance associated with double. We compared the invitro antimycobacterial activities of rifabutin and krm1648, two rifamycin derivatives, with that of rifampicin against 163 strains of mycobacterium tuberculosis. Pdf mutations in the rpob gene of mycobacterium tuberculosis. Mycobacterium tuberculosis mtb remains a major public health problem, with an effective vaccine continuing to prove elusive. To analyze characterization of the rpob gene mutations of mycobacterium tuberculosis isolated from china and to explore the association of specific mutations conferring rifampicin rif resistance with beijing genotype strains. The missing inner part of the rpob gene flanked with natural bst eii restriction sites contains an 81bp mutable region.
To identify the mutation in the rpob gene of mycobacterium tuberculosis mtb, using by sequencing method from pulmonary. Rifampin is a major drug used to treat leprosy and tuberculosis. Multidrugresistant tuberculosis mdrtb, defined as tb that is resistant to at least isoniazid inh and rifampicin rif. Most of these cases are identified only 36 months after initiation of treatment owing to incompletefailed clinical response and incomplete. An additional track containing mutations from a database of 53 known drug resistance mutations 16 aligned to the same version of the genome was used to check for mutations present in the rpob gene. Rifampicin resistance and mutation of the rpob gene in mycobacterium tuberculosis.
To determine the impact of different rpob mutations on the relative competitive fitness of rifampinresistant m. We detected the c526t and c531t mutations in the rifampicin resistancedetermining region rrdr of the rpob gene with qpcrhrm using plasmidbased controls. Progress in vaccination strategies has been hampered by a lack of appreciation of the bacteriums response to dynamic changes in the host immune environment. Rifampicin resistance mutations in the 81 bp rrdr of rpob. Quantitative polymerase chain reactionhighresolution melting qpcrhrm analysis was used to screen for mutations related to drug resistance in mycobacterium tuberculosis. Mycobacterium tuberculosis isolates cultured from 6 patients associated with an isoniazidresistant m. Shengfen wang, bing zhao, yuanyuan song, yang zhou, yu pang, xichao ou, qiang li, hui xia, yanlin zhao national tuberculosis reference laboratory, national center for tuberculosis control and prevention, chinese center for disease. For mycobacterium tuberculosis, the rifamycinresistant mutations most commonly encountered involve codons 516, 526, and 531 numbered, by convention, as in escherichia coli rpob. To illustrate this point we now report a previously uncharacterized mutation in the rpob gene of a multidrugresistant clinical isolate of m. Molecular characterization of the rpob gene mutations of. Sequence analysis of dna of mycobacterium tuberculosis lforms it was 29 mutational strains of rpob gene in 31 rifampicinresistant strains, the rate of mutation was 93.615 737 389 220 1150 1228 646 1092 1104 1111 1424 613 132 138 1329 1311 985 622 439 955 278 782 187 404 185 39 458 81 756 130 935 405 1190 1438 630 838 643 483 804 35