Detection of mutations in gyrB using denaturing high performance liquid chromatography (DHPLC) among Salmonella enterica serovar Typhi and Paratyphi A

Gupta, Ruchi, Gaind, Rajni, Chandreshwor Singh, Laishram, Paglietti, Bianca, Deb, Monorama, Rubino, Salvatore, Wain, John and Farhat Basir, Seemi (2017) Detection of mutations in gyrB using denaturing high performance liquid chromatography (DHPLC) among Salmonella enterica serovar Typhi and Paratyphi A. Transactions of the Royal Society of Tropical Medicine and Hygiene, 110 (12). pp. 684-689. ISSN 0035-9203

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Abstract

Background:- Fluoroquinolone resistance is mediated by mutations in the quinolone-resistance determining region (QRDR) of the topoisomerase genes. Denaturing high performance liquid chromatography (DHPLC) was evaluated for detection of clinically important mutations in gyrB among Salmonella. Method:- S. Typhi and S. ParatyphiA characterised for mutation in QRDR of gyrA, parC and parE were studied for mutation in gyrB by DHPLC and validated by sequencing. Result:- The DHPLC analysis was able to resolve the test mutant from isolates with wild type gyrB and distinguished mutants from other mutant by peak profile and shift in retention time. Three sequence variants were detected at codon 464, and a novel mutation Ser→Thr was also detected. gyrB mutation was associated with non classical quinolone resistance (NALS-CIPDS) in 34 isolates of S. Typhi only and was distinct from classical quinolone resistance associated with gyrA mutations (NALR-CIPDS). Conclusion: DHPLC is effective for the detection of mutation and can reduce the need forsequencing to detect clinically significant gyrB mutations..

Item Type:	Article
Uncontrolled Keywords:	decreased ciprofloxacin susceptibility,dhplc,gyrb mutation,salmonella paratyphi a,salmonella typhi
Faculty \ School:	Faculty of Medicine and Health Sciences > Norwich Medical School
UEA Research Groups:	Faculty of Medicine and Health Sciences > Research Groups > Medical Microbiology (former - to 2018)
Depositing User:	Pure Connector
Date Deposited:	13 Apr 2017 05:08
Last Modified:	22 Oct 2022 02:24
URI:	https://ueaeprints.uea.ac.uk/id/eprint/63242
DOI:	10.1093/trstmh/trx002

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