High-Throughput Typing Method To Identify a Non-Outbreak-Involved Legionella pneumophila Strain Colonizing the Entire Water Supply System in the Town of Rennes, France

Sobral, Daniel; Cann, Pierre Le; Gérard, A.; Jarraud, Sophie; Lebeau, B.; Loisy-Hamon, Fabienne; Vergnaud, Gilles; Pourcel, Christine

doi:10.1128/aem.05556-11

Cited by 26 publications

(44 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This approach would be cost-effective, since in our laboratory, the re-agents and sequencing costs for the characterization of one strain by means of recA sequence for species identification plus MLVA-10 were 36 euros compared to 112 euros for MLST analysis, with comparable hands-on time. Besides, the MLVA-10 scheme could easily be automated using fluorescent primers and a single multiplex PCR (31). Capillary electrophoresis would allow automatic size measurement of the PCR products and thus increase the accuracy of repeat number determination, as described for other bacterial species (31)(32)(33).…”

Section: Discussionmentioning

confidence: 99%

“…Besides, the MLVA-10 scheme could easily be automated using fluorescent primers and a single multiplex PCR (31). Capillary electrophoresis would allow automatic size measurement of the PCR products and thus increase the accuracy of repeat number determination, as described for other bacterial species (31)(32)(33). Furthermore, some potentially informative VNTR markers that were ruled out from the present scheme due to short repeats (Ͻ15 bp) could be included to constitute a second panel with high discriminatory power, as has been proposed for other bacteria (34).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

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

View full text Add to dashboard Cite

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...

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

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

View full text Add to dashboard Cite

show abstract

“…The MLVA used for Mycobacterium tuberculosis [9] [10]. Alternatively, the assays can be set up locally, with no need to buy kits, since all the necessary information is published [10][11][12]. MLVA is also in routine use for Streptococcus pneumoniae, with more than 1,000 genotypes publicly accessible from the the Robert Picqué Military Hospital in Bordeaux [13] and for Acinetobacter baumanii [14].…”

Section: Francementioning

confidence: 99%

“…(NA) and STTR3 (211) and different combinations of the variable loci STTR5 (11,12,13) and STTR6 (9,10), where paranthesised numbers denote allele sizes and NA (no amplification) indicates negative PCR amplification, as previously described [2].…”

Section: Denmarkmentioning

confidence: 99%

See 1 more Smart Citation

Use of multilocus variable-number tandem repeat analysis (MLVA) in eight European countries, 2012

et al. 2013

View full text Add to dashboard Cite

Genotyping of important medical or veterinary prokaryotes has become a very important tool during the last decades. Rapid development of fragment-separation and sequencing technologies has made many new genotyping strategies possible. Among these new methods is multilocus variable-number tandem repeat analysis (MLVA). Here we present an update on the use of MLVA in eight European countries (Denmark, France, Germany, Ireland, Italy, the Netherlands, Norway and Sweden). Researchers in Europe have been active in developing and implementing a large array of different assays. MLVA has been used as a typing tool in several contexts, from aiding in resolving outbreaks of food-borne bacteria to typing organisms that may pose a bioterrorist threat, as well as in scientific studies.

show abstract

Capillary electrophoresis applied to DNA: determining and harnessing sequence and structure to advance bioanalyses (2009–2014)

2015

View full text Add to dashboard Cite

This review of capillary electrophoresis methods for DNA analyses covers critical advances from 2009 to 2014, referencing 184 citations. Separation mechanisms based on free-zone capillary electrophoresis, Ogston sieving, and reptation are described. Two prevalent gel matrices for gel-facilitated sieving, which are linear polyacrylamide and polydimethylacrylamide, are compared in terms of performance, cost, viscosity, and passivation of electroosmotic flow. The role of capillary electrophoresis in the discovery, design, and characterization of DNA aptamers for molecular recognition is discussed. Expanding and emerging techniques in the field are also highlighted.

show abstract

High-Throughput Typing Method To Identify a Non-Outbreak-Involved Legionella pneumophila Strain Colonizing the Entire Water Supply System in the Town of Rennes, France

Cited by 26 publications

References 35 publications

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

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

Use of multilocus variable-number tandem repeat analysis (MLVA) in eight European countries, 2012

Capillary electrophoresis applied to DNA: determining and harnessing sequence and structure to advance bioanalyses (2009–2014)

Contact Info

Product

Resources

About