Fine-Scale Genetic Diversity among
            <i>Burkholderia pseudomallei</i>
            Soil Isolates in Northeast Thailand

U’Ren, Jana M.; Hornstra, Heidie; Pearson, Talima; Schupp, James M.; Leadem, Benjamin R.; Georgia, Shalamar; Sermswan, Rasana W.; Keim, Paul

doi:10.1128/aem.00986-07

Cited by 23 publications

(23 citation statements)

References 21 publications

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“…2A). VgrG5 sequences from Australian (MSHR668) and Southeast Asian (1026b) strains, which represent the two major clades of the species B. pseudomallei (41), and from B. mallei are nearly identical (Ն99% amino acid [aa] identity) but diverge from those of B. thailandensis and B. oklahomensis, which share 90% and 82% identity, respectively, with the 1026b sequence. Although little is known regarding virulence mechanisms, B. oklahomensis has been isolated from soil and human wound infections and is minimally pathogenic in animal models (42)(43)(44).…”

Section: Resultsmentioning

confidence: 99%

The Type VI Secretion System Spike Protein VgrG5 Mediates Membrane Fusion during Intercellular Spread by Pseudomallei Group Burkholderia Species

2014

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Pseudomallei group Burkholderia species are facultative intracellular parasites that spread efficiently from cell to cell by a mechanism involving the fusion of adjacent cell membranes. Intercellular fusion requires the function of the cluster 5 type VI secretion system (T6SS-5) and its associated valine-glycine repeat protein, VgrG5. Here we show that VgrG5 alleles are conserved and functionally interchangeable between Burkholderia pseudomallei and its relatives B. mallei, B. oklahomensis, and B. thailandensis. We also demonstrate that the integrity of the VgrG5 C-terminal domain is required for fusogenic activity, and we identify sequence motifs, including two hydrophobic segments, that are important for fusion. Mutagenesis and secretion experiments using B. pseudomallei strains engineered to express T6SS-5 in vitro show that the VgrG5 C-terminal domain is dispensable for T6SS-mediated secretion of Hcp5, demonstrating that the ability of VgrG5 to mediate membrane fusion can be uncoupled from its essential role in type VI secretion. We propose a model in which a unique fusogenic activity at the C terminus of VgrG5 facilitates intercellular spread by B. pseudomallei and related species following injection across the plasma membranes of infected cells.

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Section: Resultsmentioning

confidence: 99%

The Type VI Secretion System Spike Protein VgrG5 Mediates Membrane Fusion during Intercellular Spread by Pseudomallei Group Burkholderia Species

2014

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“…pseudomallei contains numerous VNTRs. Using a 32 VNTR system, U'Ren et al showed extensive diversity within a global B. pseudomallei isolate set (26). When 30 of these VNTR loci were used to analyze 9 epidemiologically related B. pseudomallei isolate sets, fi ne-scale diversity was found even among closely related strains, including sequential isolates from persons (13).…”

Section: Discussionmentioning

confidence: 99%

Identification of Melioidosis Outbreak by Multilocus Variable Number Tandem Repeat Analysis

Currie¹,

Haslem²,

Pearson³

et al. 2009

Emerg. Infect. Dis.

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Endemic melioidosis is caused by genetically diverse Burkholderia pseudomallei strains. However, clonal outbreaks (multiple cases caused by 1 strain) have occurred, such as from contaminated potable water. B. pseudomallei is designated a group B bioterrorism agent, which necessitates rapidly recognizing point-source outbreaks. Pulsedfi eld gel electrophoresis (PFGE) and multilocus sequence typing (MLST) can identify genetically related isolates, but results take several days to obtain. We developed a simplifi ed 4-locus multilocus variable number tandem repeat analysis (MLVA-4) for rapid typing and compared results with PFGE and MLST for a large number of well-characterized B. pseudomallei isolates. MLVA-4 compared favorably with MLST and PFGE for the same isolates; it discriminated between 65 multilocus sequence types and showed relatedness between epidemiologically linked isolates from outbreak clusters and between isolates from individual patients. MLVA-4 can establish or refute that a clonal outbreak of melioidosis has occurred within 8 hours of receipt of bacterial strains.

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“…MLVA codes can also be deduced from wholegenome sequence data (14). An MLVA scheme has been developed to type Burkholderia pseudomallei (15,16).…”

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confidence: 99%

Development of a Multiple-Locus Variable-Number Tandem-Repeat Typing Scheme for Genetic Fingerprinting of Burkholderia cenocepacia and Application to Nationwide Epidemiological Analysis

Segonds¹,

Thouverez²,

Barthe³

et al. 2015

J Clin Microbiol

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Organisms of the Burkholderia cepacia complex are especially important pathogens in cystic fibrosis (CF), with a propensity for patient-to-patient spread and long-term respiratory colonization. B. cenocepacia and Burkholderia multivorans account for the majority of infections in CF, and major epidemic clones have been recognized throughout the world. The aim of the present study was to develop and evaluate a multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) scheme for B. cenocepacia. Potential VNTR loci were identified upon analysis of the annotated genome sequences of B. cenocepacia strains AU1054, J2315, and MCO-3, and 10 of them were selected on the basis of polymorphisms and size. A collection of 100 B. cenocepacia strains, including epidemiologically related and unrelated strains, as well as representatives of the major epidemic lineages, was used to evaluate typeability, epidemiological concordance, and the discriminatory power of MLVA-10 compared with those of pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Longitudinal stability was assessed by testing 39 successive isolates from 14 patients. Typeability ranged from 0.91 to 1, except for that of one marker, which was not amplified in 53% of the B. cenocepacia IIIA strains. The MLVA types were shown to be stable in chronically colonized patients and within outbreak-related strains, with excellent epidemiological concordance. The Burkholderia cepacia complex (BCC), which includes 18 closely related species, is mainly involved in pulmonary infections in patients with cystic fibrosis (CF), although it is also occasionally recovered in nosocomial infections and/or from immunocompromised hosts. BCC infection in CF patients is associated with accelerated decline of lung function, cepacia syndrome, and poor posttransplantation outcome. The most frequently recovered species of the BCC are B. cenocepacia and Burkholderia multivorans, which have been found to be responsible for large outbreaks among CF patients, prompting the implementation of infection control measures and epidemiological surveillance. B. cenocepacia consists of four recA subgroups: IIIA, which includes the Edinburgh-Toronto electrophoretic type 12 (ET-12) epidemic lineage (in Canada and the United Kingdom) (1); IIIB, which includes the PHDC epidemic lineage (in the United States) (2) and the Midwest clone (3); IIIC, which is exclusively environmental; and IIID, which was reported to be responsible for 50% of the BCC infections in an Italian CF center (4).Putative transmissibility markers have been identified within B. cenocepacia epidemic strains. The cblA pilin gene is characteristic of ET-12 strains (5). The B. cepacia epidemic strain marker (BCESM), which is part of a pathogenicity island, was identified in ET-12 strains and in other epidemic lineages but also in unique clinical or environmental strains (6, 7), whereas the PHDC lineage is BCESM negative. Finally, the IS1363 insertion sequence was demonstrated to be characteristic of the PHDC and ET...

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Fine-Scale Genetic Diversity among Burkholderia pseudomallei Soil Isolates in Northeast Thailand

Cited by 23 publications

References 21 publications

The Type VI Secretion System Spike Protein VgrG5 Mediates Membrane Fusion during Intercellular Spread by Pseudomallei Group Burkholderia Species

The Type VI Secretion System Spike Protein VgrG5 Mediates Membrane Fusion during Intercellular Spread by Pseudomallei Group Burkholderia Species

Identification of Melioidosis Outbreak by Multilocus Variable Number Tandem Repeat Analysis

Development of a Multiple-Locus Variable-Number Tandem-Repeat Typing Scheme for Genetic Fingerprinting of Burkholderia cenocepacia and Application to Nationwide Epidemiological Analysis

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