A double mutation in the gyrA gene is necessary to produce high levels of resistance to moxifloxacin in Campylobacter spp. clinical isolates

ABSTRACT Quinolone susceptibility was analyzed in 17 clinical isolates of Corynebacterium striatum and 9 strains of Corynebacterium amycolatum by the E-test method in Mueller-Hinton agar plates. The C. striatum ATCC 6940 strain was used as a control strain. The amplified quinolone resistance determining regions of the gyrA genes of C. amycolatum and C. striatum were characterized. Four in vitro quinolone-resistant mutants of C. amycolatum were selected and analyzed. Both in vivo and in vitro quinolone-resistant strains of C. amycolatum showed high levels of fluoroquinolone resistance in strains with a double mutation leading to an amino acid change in positions 87 and 91 or positions 87 and 88 (unusual mutation) of GyrA, whereas the same concomitant mutations at amino acid positions 87 and 91 in GyrA of C. striatum produced high levels of resistance to ciprofloxacin and levofloxacin but only showed a moderate increase in the MIC of moxifloxacin, suggesting that other mechanism(s) of quinolone resistance could be involved in moxifloxacin resistance in C. amycolatum. Moreover, a PCR-RFLP-NcoI of the gyrA gene was developed to distinguish between C. amycolatum and C. striatum species.

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Association between double mutation in gyrA gene of ciprofloxacin-resistant clinical isolates of Escherichia coli and MICs.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.38.10.2477 ◽

1994 ◽

Vol 38 (10) ◽

pp. 2477-2479 ◽

Cited By ~ 127

Author(s):

J Vila ◽

J Ruiz ◽

F Marco ◽

A Barcelo ◽

P Goni ◽

...

Keyword(s):

Escherichia Coli ◽

Clinical Isolates ◽

Gyra Gene ◽

Double Mutation

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Antimicrobial resistance among Campylobacter spp. strains isolated from faecal samples of pigs chickens and ducks at Dong Thap province

Science and Technology Development Journal ◽

10.32508/stdj.v19i1.543 ◽

2016 ◽

Vol 19 (1) ◽

pp. 45-54

Author(s):

Thuy Thu Cao ◽

Hoang Van Minh Nguyen ◽

Hien Tinh Tran ◽

James Ian Campbell ◽

Stephen Baker ◽

...

Keyword(s):

23S Rrna ◽

Rrna Gene ◽

Campylobacter Coli ◽

Erythromycin Resistance ◽

Gyra Gene ◽

Pump System ◽

Double Mutation ◽

Faecal Samples ◽

23S Rrna Gene ◽

Campylobacter Spp

This study was designed to identify the genetic basis that results in the development of antibiotic resistance in Campylobacter spp. We carried out a molecular examination of 75 strains looking for resistance factors. These strains were isolated from faecal samples of pigs, chickens and ducks at Dong Thap province. There were 89.3 % strains which showed mutations on DNA gyrase-gyrA gene at the position 86 (C257T and T227G). There were also a further 32 % strains that showed mutations on gene cmeR encoding the efflux CmeABC pump system. This study has also shown that 29.3 % of Campylobacter spp. have mutations in fluoroquinolone (FQ) resistance on both genes gyrA and cmeR. In comparison with Campylobacter jejuni with the mutation C257T (Thr-86-Ile), Campylobacter coli has another mutation at T227G (Thr-86-Met). This mutation, encoding FQ resistance, is common in both swine and poultry but has not yet been identified in Viet Nam. As well as FQ resistance, resistance to erythromycine (Ery) has been detected in C. coli, in 10 strains erythromycin resistance, with a mutation at position A2075G. There was 1 strain which carried two mutations A2075G and A2074C at 23S-rRNA gene, and 2 mutant strains with a double mutation at A2075G and A2076C. In summary, there were three strains of C. coli with double mutations encoding Ery resistance at Dong Thap and this resistance characteristic has not been identified eslewhere.

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Prevalence and Antimicrobial Resistance of Campylobacter spp. Isolated from Poultry Carcasses in Poland

Journal of Food Protection ◽

10.4315/0362-028x.jfp-13-035 ◽

2013 ◽

Vol 76 (8) ◽

pp. 1451-1455 ◽

Cited By ~ 23

Author(s):

KINGA WIECZOREK ◽

IWONA KANIA ◽

JACEK OSEK

Keyword(s):

Campylobacter Jejuni ◽

23S Rrna ◽

Rrna Gene ◽

Campylobacter Coli ◽

Genetic Determinants ◽

Gyra Gene ◽

23S Rrna Gene ◽

Pcr Method ◽

Almost All ◽

Campylobacter Spp

The purpose of the present study was to determine the prevalence of Campylobacter in poultry carcasses at slaughter in Poland. For the isolated strains, resistance to selected antibiotics and the associated genetic determinants were identified. A total of 498 Campylobacter isolates were obtained from 802 poultry samples during the 2-year study period. Strains were identified to species with the PCR method; 53.6% of the strains were Campylobacter jejuni and 46.4% were Campylobacter coli. A high percentage of the tested Campylobacter strains were resistant to ciprofloxacin and nalidixic acid (74.1 and 73.5%, respectively) followed by tetracycline (47.4%) and streptomycin (20.5%). Only one C. jejuni and two C. coli isolates were resistant to gentamicin. Seventy-nine (15.9%) of the 498 strains were resistant to three or more classes of antibiotics examined. Higher levels of resistance, irrespective of the antimicrobial agent tested, were found within the C. coli group. Almost all strains resistant to quinolones (99.5%) and to tetracycline (99.6%) carried the Thr-86-to-Ile mutation in the gyrA gene and possessed the tet(O) marker, respectively. All isolates resistant to erythromycin had the A2075G mutation in the 23S rRNA gene. These results reveal that poultry carcasses in Poland are a reservoir of potentially pathogenic and antimicrobial-resistant Campylobacter strains for humans, which may pose a public health risk.

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Multiplex PCR Assay for Identifi cation and Differentiation of Campylobacter jejuni and Campylobacter coli Isolates

Folia Medica ◽

10.1515/folmed-2016-0016 ◽

2016 ◽

Vol 58 (2) ◽

pp. 95-100 ◽

Cited By ~ 3

Author(s):

Maria R. Pavlova ◽

Elina G. Dobreva ◽

Katucha I. Ivanova ◽

Galina D. Asseva ◽

Ivan N. Ivanov ◽

...

Keyword(s):

Multiplex Pcr ◽

Campylobacter Jejuni ◽

Accurate Method ◽

Clinical Isolates ◽

Campylobacter Coli ◽

Pcr Assay ◽

Biochemical Tests ◽

Multiplex Pcr Assay ◽

Hydrolysis Of ◽

Campylobacter Spp

AbstractIntroduction: Campylobacter spp. are important causative agents of gastrointestinal infections in humans. The most frequently isolated strains of this bacterial genus are Campylobacter jejuni and Campylobacter coli. To date, genetic methods for bacterial identification have not been used in Bulgaria. We optimized the multiplex PSR assay to identify Campylobacter spp. and differentiate C. jejuni from C. coli in clinical isolates. We also compared this method with the routinely used biochemical methods.Aim: To identify Campylobacter spp. and discriminate C. coli from C. jejuni in clinical isolates using multiplex PCR assay.Materials and methods: Between February 2014 and January 2015 we studied 93 stool samples taken from patients with diarrheal syndrome and identified 40 species of Campylobacter spp. in them. The clinical material was cultured in microaerophilic atmosphere, the isolated strains being biochemically diff erentiated (hydrolysis of sodium hippurate for C. jejuni, and hydrolysis of indoxyl acetate for C. coli). DNA was isolated from the strains using QiaAmp MiniKit (QIAGEN, Germany). Twenty strains were tested with multiplex PCR for the presence of these genes: cadF, characteristic for Campylobacter spp., hipO for C. jejuni and asp for C. coli.Results and discussion: The biochemical tests identified 16 strains of C. jejuni, 3 strains of C. coli, and 1 strain of C. upsaliensis. After the multiplex PCR assay the capillary gel electrophoresis confirmed 16 strains of C. jejuni, 2 strains of C. coli and 2 strains of Campylobacter spp. - because of the presence of the gene cadF. C. jejuni has the gene hipO, and it is possible that this gene may not be expressed in the biochemical differentiation yielding a negative reaction as a result. In comparison, we can conclude that the genetic differentiation is a more accurate method than the biochemical tests.Conclusion: The multiplex PCR assay is a fast, accurate method for identifi cation of Campylobacter spp. which makes it quite necessary in the clinical diagnostic practice.

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Transcriptome analysis of biofilm formation under aerobic and microaerobic conditions in clinical isolates of Campylobacter spp.

Research in Veterinary Science ◽

10.1016/j.rvsc.2021.11.009 ◽

2021 ◽

Author(s):

Elnaz Ohadi ◽

Bita Bakhshi ◽

Behrooz Sadeghi Kalani ◽

Malihe Talebi ◽

Gholamreza Irajian

Keyword(s):

Transcriptome Analysis ◽

Biofilm Formation ◽

Clinical Isolates ◽

Microaerobic Conditions ◽

Campylobacter Spp

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Comparative Analysis of Quinolone Resistance in Clinical Isolates ofKlebsiella pneumoniaeandEscherichia colifrom Chinese Children and Adults

BioMed Research International ◽

10.1155/2015/168292 ◽

2015 ◽

Vol 2015 ◽

pp. 1-6 ◽

Cited By ~ 4

Author(s):

Ying Huang ◽

James O. Ogutu ◽

Jiarui Gu ◽

Fengshu Ding ◽

Yuhong You ◽

...

Keyword(s):

Escherichia Coli ◽

Environmental Issues ◽

Quinolone Resistance ◽

Clinical Isolates ◽

Fluoroquinolone Resistance ◽

Chinese Adults ◽

Double Mutation ◽

History Of ◽

Ciprofloxacin Resistance ◽

Resistance Rates

The objective of this study was to compare quinolone resistance andgyrAmutations in clinical isolates ofKlebsiella pneumoniaeandEscherichia colifrom Chinese adults who used quinolone in the preceding month and children without any known history of quinolone administration. The antimicrobial susceptibilities of 61 isolates from children and 79 isolates from adults were determined. The mutations in the quinolone resistance-determining regions ingyrAgene were detected by PCR and DNA sequencing. Fluoroquinolone resistance and types ofgyrAmutations in isolates from children and adults were compared and statistically analyzed. No significant differences were detected in the resistance rates of ciprofloxacin and levofloxacin between children and adults among isolates of the two species (allP>0.05). The double mutation Ser83→Leu + Asp87→Asn in the ciprofloxacin-resistant isolates occurred in 73.7% isolates from the children and 67.9% from the adults, respectively (P=0.5444). Children with no known history of quinolone administration were found to carry fluoroquinolone-resistantEnterobacteriaceaeisolates. The occurrence of ciprofloxacin resistance and the major types ofgyrAmutations in the isolates from the children were similar to those from adults. The results indicate that precautions should be taken on environmental issues resulting from widespread transmission of quinolone resistance.

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