Molecular characterization of Coxiella burnetii isolates by infrequent restriction site-PCR and MLVA typing View Full Text


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Article Info

DATE

2006-04-26

AUTHORS

Nathalie Arricau-Bouvery, Yolande Hauck, Awatef Bejaoui, Dimitrios Frangoulidis, Christelle C Bodier, Armel Souriau, Hermann Meyer, Heinrich Neubauer, Annie Rodolakis, Gilles Vergnaud

ABSTRACT

BACKGROUND: Coxiella burnetii, the causative agent of Q fever, has a wide host range. Few epidemiological tools are available, and they are often expensive or not easily standardized across laboratories. In this work, C. burnetii isolates from livestock and ticks were typed using infrequent restriction site-PCR (IRS-PCR) and multiple loci variable number of tandem repeats (VNTR) analysis (MLVA). RESULTS: By applying IRS-PCR, 14 C. burnetii isolates could be divided into six groups containing up to five different isolates. Clustering as deduced from MLVA typing with 17 markers provided an increased resolution with an excellent agreement to IRS-PCR, and with the plasmid type of each strain. MLVA was then applied to 28 additional C. burnetii isolates of different origin and 36 different genotypes were identified among the 42 isolates investigated. The clustering obtained is in agreement with published Multiple Locus Sequence Typing (MLST) data. Two panels of markers are proposed, panel 1 which can be confidently typed on agarose gel at a lower cost and in any laboratory setting (10 minisatellite markers with a repeat unit larger than 9 bp), and panel 2 which comprises 7 microsatellites and provides a higher discriminatory power. CONCLUSION: Our analyses demonstrate that MLVA is a powerful and promising molecular typing tool with a high resolution and of low costs. The consistency of the results with independent methods suggests that MLVA can be applied for epidemiological studies. The resulting data can be queried on a dedicated MLVA genotyping Web service. More... »

PAGES

38-38

References to SciGraph publications

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  • 2004-06-08. Tandem repeats analysis for the high resolution phylogenetic analysis of Yersinia pestis in BMC MICROBIOLOGY
  • 2006-02-09. Evaluation and selection of tandem repeat loci for a Brucella MLVA typing assay in BMC MICROBIOLOGY
  • 2005-11-16. Evaluation and selection of tandem repeat loci for Streptococcus pneumoniae MLVA strain typing in BMC MICROBIOLOGY
  • 1993-07. Polymorphism in DNA restriction patterns of Coxiella burnetii isolates investigated by pulsed field gel electrophoresis and image analysis in EUROPEAN JOURNAL OF EPIDEMIOLOGY
  • 1993-07. Plasmid based differentiation and detection of Coxiella burnetii in clinical samples in EUROPEAN JOURNAL OF EPIDEMIOLOGY
  • 2001-03-30. A tandem repeats database for bacterial genomes: application to the genotyping of Yersinia pestis and Bacillus anthracis in BMC MICROBIOLOGY
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    http://scigraph.springernature.com/pub.10.1186/1471-2180-6-38

    DOI

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    DIMENSIONS

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    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/16640773


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    36 schema:description BACKGROUND: Coxiella burnetii, the causative agent of Q fever, has a wide host range. Few epidemiological tools are available, and they are often expensive or not easily standardized across laboratories. In this work, C. burnetii isolates from livestock and ticks were typed using infrequent restriction site-PCR (IRS-PCR) and multiple loci variable number of tandem repeats (VNTR) analysis (MLVA). RESULTS: By applying IRS-PCR, 14 C. burnetii isolates could be divided into six groups containing up to five different isolates. Clustering as deduced from MLVA typing with 17 markers provided an increased resolution with an excellent agreement to IRS-PCR, and with the plasmid type of each strain. MLVA was then applied to 28 additional C. burnetii isolates of different origin and 36 different genotypes were identified among the 42 isolates investigated. The clustering obtained is in agreement with published Multiple Locus Sequence Typing (MLST) data. Two panels of markers are proposed, panel 1 which can be confidently typed on agarose gel at a lower cost and in any laboratory setting (10 minisatellite markers with a repeat unit larger than 9 bp), and panel 2 which comprises 7 microsatellites and provides a higher discriminatory power. CONCLUSION: Our analyses demonstrate that MLVA is a powerful and promising molecular typing tool with a high resolution and of low costs. The consistency of the results with independent methods suggests that MLVA can be applied for epidemiological studies. The resulting data can be queried on a dedicated MLVA genotyping Web service.
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    44 Locus Sequence Typing (MLST) data
    45 MLVA
    46 Multiple Locus Sequence Typing (MLST) data
    47 Panel 1
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    49 Sequence Typing (MLST) data
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