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A new highly discriminatory multiplex capillary-based MLVA assay as a tool for the epidemiological survey of Pseudomonas aeruginosa in cystic fibrosis patients

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Abstract

Multiple locus variable number of tandem repeats (VNTR) analysis (MLVA) has been shown to provide a high level of information for epidemiological investigations and the follow-up of Pseudomonas aeruginosa chronic infection. In the present study, an automatized MLVA assay has been developed for the analysis of 16 VNTRs in two multiplex polymerase chain reactions (PCRs), followed by capillary electrophoresis. The result in the form of a code is directly usable for clustering analyses. This MLVA-16Orsay scheme was applied to the genotyping of 83 isolates from eight cystic fibrosis patients, demonstrating that the same genotype persisted during eight years of chronic infection in the majority of cases. Comparison with pulsed-field gel electrophoresis (PFGE) analysis showed that both methods were congruent, MLVA providing, in some cases, additional informativity. The evolution of strains during long-term infection was revealed by the presence of VNTR variants.

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Acknowledgments

D.S., F.L.-H., and B.L. are employees of Ceeram and hold stocks. This study was performed with the support of the association Vaincre La Mucoviscidose (grant no. RC0630). The development of tools for the surveillance of bacterial pathogens is supported by the French Direction Générale de l’Armement.

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Authors and Affiliations

  1. Institut de Génétique et Microbiologie, University Paris-Sud, UMR 8621, 91405, Orsay, France

    D. Sobral, K. Hormigos, G. Vergnaud & C. Pourcel

  2. CNRS, 91405, Orsay, France

    D. Sobral, K. Hormigos, G. Vergnaud & C. Pourcel

  3. Ceeram (Centre Européen d’Expertise et de Recherche sur les Agents Microbiens), Allée de la Filée, 44244, La Chapelle-sur-Erdre, France

    D. Sobral, B. Lebeau & F. Loisy-Hamon

  4. Laboratoire de Microbiologie, Hôpital Robert Debré, Université Paris 7, Assistance Publique—Hôpitaux de Paris (APHP), 75019, Paris, France

    P. Mariani-Kurkdjian & E. Bingen

  5. Laboratoire de Microbiologie, Hôpital Armand Trousseau, Université Paris 6, Assistance Publique—Hôpitaux de Paris (APHP), 75012, Paris, France

    H. Vu-Thien

  6. Département de Gastroentérologie Pédiatrique, Nutrition et Mucoviscidose, Université Paris 7, Hôpital Robert Debré, Assistance Publique—Hôpitaux de Paris (APHP), 75019, Paris, France

    A. Munck

  7. DGA/MRIS—Mission pour la Recherche et l’Innovation Scientifique, 92221, Bagneux, France

    G. Vergnaud

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  1. D. Sobral
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  2. P. Mariani-Kurkdjian
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  3. E. Bingen
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  4. H. Vu-Thien
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  7. F. Loisy-Hamon
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  8. A. Munck
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  9. G. Vergnaud
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  10. C. Pourcel
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Corresponding author

Correspondence to C. Pourcel.

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Sobral, D., Mariani-Kurkdjian, P., Bingen, E. et al. A new highly discriminatory multiplex capillary-based MLVA assay as a tool for the epidemiological survey of Pseudomonas aeruginosa in cystic fibrosis patients. Eur J Clin Microbiol Infect Dis 31, 2247–2256 (2012). https://doi.org/10.1007/s10096-012-1562-5

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  • DOI: https://doi.org/10.1007/s10096-012-1562-5

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