Geographical distribution of Brucella melitensis inferred from rpoB gene variation

Tan, Kian Lam; Tan, Yung-Chie; Chang, Li-Yen; Lee, Kok Wei; Nor'e, Siti Sarah; Yee, Wai-Yan; Hoh, Chee-Choong; AbuBakar, Sazaly

doi:10.3855/jidc.7598

Cited by 4 publications

(3 citation statements)

References 15 publications

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“…Data on these mutations can be used as markers in the development of new systems for identifying the geographical region of B. melitensis strains origin. In 2017, Kim-Kee Tan et al published the results of the SNP analysis of the nucleotide sequence of the rpoB gene to determine the origin of B. melitensis strains [ 34 ]. In the long term, this approach can be adapted to study representatives of other epidemic-significant Brucella species.…”

Section: Discussionmentioning

confidence: 99%

Global evolution and phylogeography of Brucella melitensis strains

et al. 2018

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BackgroundBrucellosis is a bacterial zoonotic disease. Annually in the world more than 500,000 new cases of brucellosis in humans are registered. In this study, we propose an evolutionary model of the historical distribution of B. melitensis based on the full-genomic SNP analysis of 98 strains.ResultsWe performed an analysis of the SNP of the complete genomes of 98 B. melitensis strains isolated in different geographical regions of the world to obtain relevant information on the population structure, genetic diversity and the evolution history of the species. Using genomic sequences of 21 strains of B. melitensis isolated in Russia and WGS data from the NCBI database, it was possible to identify five main genotypes and 13 species genotypes for analysis. Data analysis based on the Bayesian Phylogenetics and Phylogeography method allowed to determine the regions of geographical origin and the expected pathways of distribution of the main lines (genotypes and subgenotypes) of the pathogen.ConclusionsWithin the framework of our study, the model of global evolution and phylogeography of B. melitensis strains isolated in various regions of the planet was proposed for the first time. The sets of unique specific SNPs described in our study, for all identified genotypes and subgenotypes, can be used to develop new bacterial typing and identification systems for B. melitensis.Electronic supplementary materialThe online version of this article (10.1186/s12864-018-4762-2) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Global evolution and phylogeography of Brucella melitensis strains

et al. 2018

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“…Several PCR-based methods have been used to determine the exact molecular biomarkers to determine Brucella molecular type (12). Various studies have reported that the use of the RNA polymerase (rpoB) β subunit gene, which is very suitable for phylogenetic analysis and identification Brucella strains, especially in highly homologous isolates (8,15,16 RpoB-based genotyping also allows the identification of new bacterial species and analysis of the bacterial community (16). It can also describe rpoB gene mutations that play a very important role in rifampicin resistance (17).…”

Section: Introductionmentioning

confidence: 99%

Genetic Diversity and Phylogenetic Relationship of Clinical Isolates of Brucella melitensis Based on Gene Polymorphism of β Subunit of RNA Polymerase (rpoB) Gene in Iran

Bazrgari

Garosi

Dadar

2020

Iran J Med Microbiol

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Background: The prevalence of Brucella infections in animals and humans has indicated the important need for different regional/local reference laboratories to use valid species-determining approaches to facilitate and compare data exchange. The purpose of current study was to evaluate the RNA Polymerase Beta Subunit (rpoB) as a molecular marker in Brucella species differentiation and to determine the genotype of Brucella melitensis species using single-nucleotide polymorphism (SNP) analysis. Materials & Methods: In this study, blood and cerebrospinal fluid (CSF) samples were taken from 108 patients with brucellosis. After culturing the samples insupplemented Brucella agar, eleven isolates of Brucella bacteria were isolated and identified by classical and molecular biotyping methods. Then the complete sequence of their rpoB gene was multiplied and sequenced. Sequencing results were analyzed by Mega6 program. Results: According to the results, the rpoB gene was able to differentiate between Brucella species and other bacteria.Moreover, the rpoB typing grouped the majority of Iranian isolates in the rpoB type 2, while only one strain belonged to the rpoB type 1. Among the 10 isolates of rpoB type 2, there are six different isolates with only one unique type-2 SNPs in codon 985, which gives rise to new genotype 2 variants. Conclusion: Our results shown a high discriminative power of rpoB gene among B. melitensis strains from some regions of Iran, which leads to accurate genotype and identification of these bacteria.

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“…On the other hand, the use of the DNA-directed RNA polymerase subunit beta (rpoB ) gene, expressing the b-subunit of RNA polymerase, appeared to be highly effective for detecting the rpoB mutations causing rifampin resistance (15). The evaluation of the rifampin susceptibility profiles of B. melitensis and B. abortus is critical as this antibiotic is the most widely recommended and applied therapeutic agent for the treatment of human brucellosis (16,17). Resistance to rifampin is a growing problem in both developed and developing countries (18)(19)(20).…”

Section: Introductionmentioning

confidence: 99%

Investigation of Mutations in the Rifampin-Resistance-Determining Region of the rpoB Gene of Brucella melitensis by Gene Analysis

Dadar

Bazrgari

Garosi

et al. 2021

Jundishapur J Microbiol

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Background: RNA polymerase beta subunit (rpoB) gene analysis in bacterial communities is known as a method for determining rifampin sensitivity and genetic diversity among Brucella spp. Detection of antibiotic resistance among Brucella isolates can be a critical approach to control brucellosis. However, rpoB gene analysis of Brucella melitensis for assessing rifampicin resistance has not yet been performed in Iran, which is considered an endemic area for brucellosis. Objectives: The aim of this study was to analyze the whole sequence of rpoB genes of different B. melitensis isolates from humans to identify the single-nucleotide polymorphisms (SNPs) and mutations related to rifampin resistance and to analyze the genetic diversity of these bacteria in Iran. Methods: Between 2017 and 2019, a total of 156 blood samples along with 12 synovial fluid specimens were collected from brucellosis patients in different Iranian provinces and subjected to bacterial culture in Brucella selective media. Brucella identification was carried out using classical biotyping and molecular examinations. Polymerase chain reaction (PCR)-based amplification of the rpoB gene was performed by specific rpoB primers for whole gene sequencing. The antimicrobial susceptibility of Brucella isolates was assessed using disk diffusion susceptibility tests and minimal inhibitory concentration (MIC) methods. The presence of rifampin-binding sites and SNPs were investigated through rpoB whole gene sequencing. Results: Clinical B. melitensis isolates were obtained from blood (13) and synovial fluid (1) samples of patients from different regions of Iran. The results of MIC and disk diffusion susceptibility tests showed that all the isolates were sensitive to rifampin except for one isolate showing intermediate rifampin resistance based on the standards defined for slow-growing bacteria by the Clinical and Laboratory Standards Institute (CLSI). Gene analysis for identifying the mutations related to rifampin resistance and investigating genetic diversity showed that none of the B. melitensis isolates had missense mutations, confirming the susceptibility of all the studied isolates to rifampin. Conclusions: The present study revealed that rpoB gene analysis could be used for the efficient and precise identifying of the mutations related to rifampin resistance, investigating rifampin binding sites, and genotyping Brucella species. Furthermore, the identification of B. melitensis isolates with intermediate resistance to rifampicin highlighted the importance of periodically carrying out antibiotic susceptibility testing. The molecular detection of rpoB mutations in different Brucella isolates may help to prevent the spread of rifampin-resistant Brucella spp. among humans and livestock.

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Geographical distribution of Brucella melitensis inferred from rpoB gene variation

Cited by 4 publications

References 15 publications

Global evolution and phylogeography of Brucella melitensis strains

Global evolution and phylogeography of Brucella melitensis strains

Genetic Diversity and Phylogenetic Relationship of Clinical Isolates of Brucella melitensis Based on Gene Polymorphism of β Subunit of RNA Polymerase (rpoB) Gene in Iran

Investigation of Mutations in the Rifampin-Resistance-Determining Region of the rpoB Gene of Brucella melitensis by Gene Analysis

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