Molecular Characterization of Extraintestinal Escherichia coli Isolates in Japan: Relationship between Sequence Types and Mutation Patterns of Quinolone Resistance-Determining Regions Analyzed by Pyrosequencing
Nozomi Aoike,Tomoo Saga,Ryuji Sakata,Ayumi Yoshizumi,Soichiro Kimura,Morihiro Iwata,Sadako Yoshizawa,Yasuyuki Sugasawa,Yoshikazu Ishii,Keizo Yamaguchi,Kazuhiro Tateda +10 more
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TLDR
A pyrosequencing-based high-throughput method for analyzing the nucleotide sequence of the quinolone resistance-determining regions (QRDRs) of gyrA and parC successfully determined the QRDR sequences of 139 out of 140 clinical Escherichia coli isolates, 28% of which were nonsusceptible to ciprofloxacin.Abstract:
Infection from fluoroquinolone-resistant Enterobacteriaceae is an increasing health problem worldwide. In the present study, we developed a pyrosequencing-based high-throughput method for analyzing the nucleotide sequence of the quinolone resistance-determining regions (QRDRs) of gyrA and parC. By using this method, we successfully determined the QRDR sequences of 139 out of 140 clinical Escherichia coli isolates, 28% of which were nonsusceptible to ciprofloxacin. Sequence results obtained by the pyrosequencing method were in complete agreement with those obtained by the Sanger method. All fluoroquinolone-resistant isolates (n = 35; 25%) contained mutations leading to three or four amino acid substitutions in the QRDRs. In contrast, all isolates lacking a mutation in the QRDR (n = 81; 57%) were susceptible to ciprofloxacin, levofloxacin, and nalidixic acid. The qnr determinants, namely, the qnrA, qnrB, and qnrS genes, were not detected in the isolates, and the aac(6')-Ib-cr gene was detected in 2 (1.4%) of the isolates. Multilocus sequence typing of 34 randomly selected isolates revealed that sequence type 131 (ST131) (n = 7; 20%) is the most prevalent lineage and is significantly resistant to quinolones (P < 0.01). The genetic background of quinolone-susceptible isolates seemed more diverse, and interestingly, neighboring STs of ST131 in the phylogenetic tree were all susceptible to ciprofloxacin. In conclusion, our investigation reveals the relationship between fluoroquinolone resistance caused by mutations of QRDRs and the population structure of clinical extraintestinal E. coli isolates. This high-throughput method for analyzing QRDR mutations by pyrosequencing is a powerful tool for epidemiological studies of fluoroquinolone resistance in bacteria.read more
Citations
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Journal ArticleDOI
Global Extraintestinal Pathogenic Escherichia coli (ExPEC) Lineages.
TL;DR: Systematic, unbiased tracking of predominant ExPEC lineages will direct research toward better treatment and prevention strategies for extraintestinal infections.
Journal ArticleDOI
Prevention and control of multi-drug-resistant Gram-negative bacteria: recommendations from a Joint Working Party.
Apr Wilson,David M. Livermore,Jonathan A. Otter,R. E. Warren,P.J. Jenks,David A Enoch,W. Newsholme,B. Oppenheim,Alistair Leanord,Cliodna A. M. McNulty,G. Tanner,Sarah D. Bennett,M. Cann,Jennifer Bostock,E. Collins,S. Peckitt,L. Ritchie,Carole Fry,Peter M. Hawkey +18 more
TL;DR: Recommendations are made in the following areas: screening, diagnosis and infection control precautions including hand hygiene, single-room accommodation, and environmental screening and cleaning, and where there are species differences.
Journal ArticleDOI
Whole-Genome Analysis of Antimicrobial-Resistant and Extraintestinal Pathogenic Escherichia coli in River Water
Ryota Gomi,Tomonari Matsuda,Yasufumi Matsumura,Masaki Yamamoto,Michio Tanaka,Satoshi Ichiyama,Minoru Yoneda +6 more
TL;DR: The results indicate contamination of surface waters by E. coli strains belonging to clinically important clonal groups, demonstrating the clonal distribution of environmental ExPEC strains.
Journal ArticleDOI
Double-Serine Fluoroquinolone Resistance Mutations Advance Major International Clones and Lineages of Various Multi-Drug Resistant Bacteria.
TL;DR: The paper shows that favorable fitness in the major STs/lineages of these pathogens was principally attained by the capacity of evolving mutations in the fluoroquinolone-binding serine residues of both the DNA gyrase and topoisomerase IV enzymes.
Journal ArticleDOI
Occurrence of Clinically Important Lineages, Including the Sequence Type 131 C1-M27 Subclone, among Extended-Spectrum-β-Lactamase-Producing Escherichia coli in Wastewater
Ryota Gomi,Tomonari Matsuda,Yasufumi Matsumura,Masaki Yamamoto,Michio Tanaka,Satoshi Ichiyama,Minoru Yoneda +6 more
TL;DR: It is indicated that ESBLEC strains belonging to clinically important lineages, including the C1-M27 clade, may disseminate into the environment through wastewater, highlighting the need to monitor for antibiotic resistance in wastewater.
References
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Increasing extended-spectrum β-lactamase production and quinolone resistance among Gram-negative bacilli causing intra-abdominal infections in the Asia/Pacific region: Data from the Smart Study 2002–2006
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