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, YajunRoumagnac, 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.Issue Date
2010-08MeSH
Anti-Bacterial AgentsDNA Gyrase
DNA Topoisomerase IV
DNA, Bacterial
Fluoroquinolones
Genotype
Humans
Microbial Sensitivity Tests
Mutation, Missense
Polymorphism, Single Nucleotide
Salmonella paratyphi A
Salmonella typhi
Metadata
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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 chemotherapyDOI
10.1093/jac/dkq175PubMed ID
20511368Abstract
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
ArticleLanguage
enISSN
1460-2091ae974a485f413a2113503eed53cd6c53
10.1093/jac/dkq175
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