Molecular typing and profiling of topoisomerase mutations causing resistance to ciprofloxacin and levofloxacin in Elizabethkingia species

Objectives Several Elizabethkingia species often exhibit extensive antibiotic resistance, causing infections associated with severe morbidity and high mortality rates worldwide. In this study, we determined fluoroquinolone susceptibility profiles of clinical Elizabethkingia spp. isolates and investi...

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Main Authors: Ming-Jr Jian, Yun-Hsiang Cheng, Cherng-Lih Perng, Hung-Sheng Shang
Format: Article
Language:English
Published: PeerJ Inc. 2018-09-01
Series:PeerJ
Subjects:
Online Access:https://peerj.com/articles/5608.pdf
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author Ming-Jr Jian
Yun-Hsiang Cheng
Cherng-Lih Perng
Hung-Sheng Shang
author_facet Ming-Jr Jian
Yun-Hsiang Cheng
Cherng-Lih Perng
Hung-Sheng Shang
author_sort Ming-Jr Jian
collection DOAJ
description Objectives Several Elizabethkingia species often exhibit extensive antibiotic resistance, causing infections associated with severe morbidity and high mortality rates worldwide. In this study, we determined fluoroquinolone susceptibility profiles of clinical Elizabethkingia spp. isolates and investigated the resistance mechanisms. Methods In 2017–2018, 131 Elizabethkingia spp. isolates were recovered from specimens collected at tertiary care centers in northern Taiwan. Initial species identification using the Vitek MS system and subsequent verification by 16S rRNA sequencing confirmed the presence of Elizabethkingia anophelis (n = 111), E. miricola (n = 11), and E. meningoseptica (n = 9). Fluoroquinolone susceptibility was determined using the microbroth dilution method, and fluoroquinolone resistance genes were analyzed by sequencing. Results Among Elizabethkingia spp. isolates, 91% and 77% were resistant to ciprofloxacin and levofloxacin, respectively. The most prevalent alterations were two single mutations in GyrA, Ser83Ile, and Ser83Arg, detected in 76% of the isolates exhibiting fluoroquinolone MIC between 8 and 128 μg/ml. Another GyrA single mutation, Asp87Asn, was identified in two quinolone-resistant E. miricola strains. None of the isolates had alterations in GyrB, ParC, or ParE. We developed a high-resolution melting assay for rapid identification of the prevalent gyrA gene mutations. The genetic relationship between the isolates was evaluated by random amplified polymorphic DNA PCR that yielded diverse pulsotypes, indicating the absence of any temporal or spatial overlap among the patients during hospitalization. Conclusion Our analysis of fluoroquinolone-resistant Elizabethkingia spp. isolates provides information for further research on the variations of the resistance mechanism and potential clinical guidance for infection management.
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spelling doaj.art-6c4fb215b45548a5947462d7a93524272023-12-03T00:47:36ZengPeerJ Inc.PeerJ2167-83592018-09-016e560810.7717/peerj.5608Molecular typing and profiling of topoisomerase mutations causing resistance to ciprofloxacin and levofloxacin in Elizabethkingia speciesMing-Jr Jian0Yun-Hsiang Cheng1Cherng-Lih Perng2Hung-Sheng Shang3Graduate Institute of Medical Science, National Defense Medical Center, Taipei, TaiwanGraduate Institute of Medical Science, National Defense Medical Center, Taipei, TaiwanGraduate Institute of Medical Science, National Defense Medical Center, Taipei, TaiwanGraduate Institute of Medical Science, National Defense Medical Center, Taipei, TaiwanObjectives Several Elizabethkingia species often exhibit extensive antibiotic resistance, causing infections associated with severe morbidity and high mortality rates worldwide. In this study, we determined fluoroquinolone susceptibility profiles of clinical Elizabethkingia spp. isolates and investigated the resistance mechanisms. Methods In 2017–2018, 131 Elizabethkingia spp. isolates were recovered from specimens collected at tertiary care centers in northern Taiwan. Initial species identification using the Vitek MS system and subsequent verification by 16S rRNA sequencing confirmed the presence of Elizabethkingia anophelis (n = 111), E. miricola (n = 11), and E. meningoseptica (n = 9). Fluoroquinolone susceptibility was determined using the microbroth dilution method, and fluoroquinolone resistance genes were analyzed by sequencing. Results Among Elizabethkingia spp. isolates, 91% and 77% were resistant to ciprofloxacin and levofloxacin, respectively. The most prevalent alterations were two single mutations in GyrA, Ser83Ile, and Ser83Arg, detected in 76% of the isolates exhibiting fluoroquinolone MIC between 8 and 128 μg/ml. Another GyrA single mutation, Asp87Asn, was identified in two quinolone-resistant E. miricola strains. None of the isolates had alterations in GyrB, ParC, or ParE. We developed a high-resolution melting assay for rapid identification of the prevalent gyrA gene mutations. The genetic relationship between the isolates was evaluated by random amplified polymorphic DNA PCR that yielded diverse pulsotypes, indicating the absence of any temporal or spatial overlap among the patients during hospitalization. Conclusion Our analysis of fluoroquinolone-resistant Elizabethkingia spp. isolates provides information for further research on the variations of the resistance mechanism and potential clinical guidance for infection management.https://peerj.com/articles/5608.pdfElizabethkingia spp.Fluoroquinolone resistancegyrAMolecular typingHigh-resolution melting
spellingShingle Ming-Jr Jian
Yun-Hsiang Cheng
Cherng-Lih Perng
Hung-Sheng Shang
Molecular typing and profiling of topoisomerase mutations causing resistance to ciprofloxacin and levofloxacin in Elizabethkingia species
PeerJ
Elizabethkingia spp.
Fluoroquinolone resistance
gyrA
Molecular typing
High-resolution melting
title Molecular typing and profiling of topoisomerase mutations causing resistance to ciprofloxacin and levofloxacin in Elizabethkingia species
title_full Molecular typing and profiling of topoisomerase mutations causing resistance to ciprofloxacin and levofloxacin in Elizabethkingia species
title_fullStr Molecular typing and profiling of topoisomerase mutations causing resistance to ciprofloxacin and levofloxacin in Elizabethkingia species
title_full_unstemmed Molecular typing and profiling of topoisomerase mutations causing resistance to ciprofloxacin and levofloxacin in Elizabethkingia species
title_short Molecular typing and profiling of topoisomerase mutations causing resistance to ciprofloxacin and levofloxacin in Elizabethkingia species
title_sort molecular typing and profiling of topoisomerase mutations causing resistance to ciprofloxacin and levofloxacin in elizabethkingia species
topic Elizabethkingia spp.
Fluoroquinolone resistance
gyrA
Molecular typing
High-resolution melting
url https://peerj.com/articles/5608.pdf
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