MPI für Infektionsbiologie / Department of Molecular Biology |
|A multiplex single nucleotide polymorphism typing assay for detecting mutations that result in decreased fluoroquinolone susceptibility in Salmonella enterica serovars Typhi and Paratyphi A|
|Authors:||Song, Yajun; Roumagnac, Philippe; Weill, François-Xavier; Wain, John; Dolecek, Christiane; Mazzoni, Camila J.; Holt, Kathryn E.; Achtman, Mark|
|Date of Publication (YYYY-MM-DD):||2010-08|
|Title of Journal:||Journal of Antimicrobial Chemotherapy|
|Journal Abbrev.:||J. Antimicrob. Chemother.|
|Issue / Number:||8|
|Copyright:||© The Author 2010. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
|Abstract / Description:||Decreased susceptibility to fluoroquinolones has become a major problem for the successful therapy of human infections caused by Salmonella enterica, especially the life-threatening typhoid and paratyphoid fevers. By using Luminex xTAG beads, we developed a rapid, reliable and cost-effective multiplexed genotyping assay for simultaneously detecting 11 mutations in gyrA, gyrB and parE of S. enterica serovars Typhi and Paratyphi A that result in nalidixic acid resistance (Nal(R)) and/or decreased susceptibility to fluoroquinolones. This assay yielded unambiguous single nucleotide polymorphism calls on extracted DNA from 292 isolates of Salmonella Typhi (Nal(R) = 223 and Nal(S) = 69) and 106 isolates of Salmonella Paratyphi A (Nal(R) = 24 and Nal(S) = 82). All of the 247 Nal(R) Salmonella Typhi and Salmonella Paratyphi A isolates were found to harbour at least one of the target mutations, with GyrA Phe-83 as the most common one (143/223 for Salmonella Typhi and 18/24 for Salmonella Paratyphi A). We also identified three GyrB mutations in eight Nal(S) Salmonella Typhi isolates (six for GyrB Phe-464, one for GyrB Leu-465 and one for GyrB Asp-466), and mutations GyrB Phe-464 and GyrB Asp-466 seem to be related to the decreased ciprofloxacin susceptibility phenotype in Salmonella Typhi. This assay can also be used directly on boiled single colonies. The assay presented here would be useful for clinical and reference laboratories to rapidly screen quinolone-resistant isolates of Salmonella Typhi and Salmonella Paratyphi A, and decipher the underlying genetic changes for epidemiological purposes.|
|Free Keywords:||genotyping; DNA gyrase; mechanisms of resistance; Salmonella|
|Comment of the Author/Creator:||This work was supported by Science Foundation Ireland (05/FE1/B882 to M. A.) and The Wellcome Trust (to K. E. H. via her doctoral supervisor, Dr Gordon Dougan).|
|External Publication Status:||published|
|Communicated by:||Hilmar Fünning|
|Affiliations:||MPI für Infektionsbiologie/Department of Molecular Biology|
|External Affiliations:||Univ Coll Cork, Environm Res Inst, Cork, Ireland.; Ctr Cooperat Int Rech Agron Dev, UMR BGPI, F-34398 Montpellier 5, France.; Inst Pasteur, Lab Bacteries Pathogenes Enter, Paris, France.; HPA Ctr Infect, Lab Gastrointestinal Pathogens, London NW9 5EQ, England.; Wellcome Trust Sanger Inst, Cambridge CB10 1SA, England.; Hosp Trop Dis, Oxford Univ Clin Res Unit, Wellcome Trust Major Overseas Programme, Thanh Pho Ho Chi Minh, Vietnam.|
|Identifiers:||ISI:000280018400016 [ID No:1] |
ISSN:0305-7453 [ID No:2]
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