A multiplex single nucleotide polymorphism typing assay for detecting mutations that result in decreased fluoroquinolone susceptibility in Salmonella enterica serovars Typhi and Paratyphi A.

Hdl Handle:
http://hdl.handle.net/10147/113885
Title:
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
Affiliation:
Environmental Research Institute, University College Cork, Lee Road, Cork, Ireland.
Citation:
A multiplex single nucleotide polymorphism typing assay for detecting mutations that result in decreased fluoroquinolone susceptibility in Salmonella enterica serovars Typhi and Paratyphi A. 2010, 65 (8):1631-41 J. Antimicrob. Chemother.
Journal:
The Journal of antimicrobial chemotherapy
Issue Date:
Aug-2010
URI:
http://hdl.handle.net/10147/113885
DOI:
10.1093/jac/dkq175
PubMed ID:
20511368
Abstract:
OBJECTIVES: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
Item Type:
Article
Language:
en
MeSH:
Anti-Bacterial Agents; DNA Gyrase; DNA Topoisomerase IV; DNA, Bacterial; Fluoroquinolones; Genotype; Humans; Microbial Sensitivity Tests; Mutation, Missense; Polymorphism, Single Nucleotide; Salmonella paratyphi A; Salmonella typhi
ISSN:
1460-2091

Full metadata record

DC FieldValue Language
dc.contributor.authorSong, Yajunen
dc.contributor.authorRoumagnac, Philippeen
dc.contributor.authorWeill, François-Xavieren
dc.contributor.authorWain, Johnen
dc.contributor.authorDolecek, Christianeen
dc.contributor.authorMazzoni, Camila Jen
dc.contributor.authorHolt, Kathryn Een
dc.contributor.authorAchtman, Marken
dc.date.accessioned2010-10-27T07:58:27Z-
dc.date.available2010-10-27T07:58:27Z-
dc.date.issued2010-08-
dc.identifier.citationA multiplex single nucleotide polymorphism typing assay for detecting mutations that result in decreased fluoroquinolone susceptibility in Salmonella enterica serovars Typhi and Paratyphi A. 2010, 65 (8):1631-41 J. Antimicrob. Chemother.en
dc.identifier.issn1460-2091-
dc.identifier.pmid20511368-
dc.identifier.doi10.1093/jac/dkq175-
dc.identifier.urihttp://hdl.handle.net/10147/113885-
dc.description.abstractOBJECTIVES: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.-
dc.language.isoenen
dc.subject.meshAnti-Bacterial Agents-
dc.subject.meshDNA Gyrase-
dc.subject.meshDNA Topoisomerase IV-
dc.subject.meshDNA, Bacterial-
dc.subject.meshFluoroquinolones-
dc.subject.meshGenotype-
dc.subject.meshHumans-
dc.subject.meshMicrobial Sensitivity Tests-
dc.subject.meshMutation, Missense-
dc.subject.meshPolymorphism, Single Nucleotide-
dc.subject.meshSalmonella paratyphi A-
dc.subject.meshSalmonella typhi-
dc.titleA multiplex single nucleotide polymorphism typing assay for detecting mutations that result in decreased fluoroquinolone susceptibility in Salmonella enterica serovars Typhi and Paratyphi A.en
dc.typeArticleen
dc.contributor.departmentEnvironmental Research Institute, University College Cork, Lee Road, Cork, Ireland.en
dc.identifier.journalThe Journal of antimicrobial chemotherapyen
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