Increased Quinolone-Resistant Mutations of gyrA and parC Genes after Pouchitis Treatment with Ciprofloxacin

2020 ◽  
Vol 37 (4) ◽  
pp. 321-330
Author(s):  
Kouhei Fukushima ◽  
Takashi Saito ◽  
Atsushi Kohyama ◽  
Kazuhiro Watanabe
Keyword(s):  
Resistance Mutation ◽  
Mutation Rates ◽  
Pcr Cloning ◽  
Gyra Gene ◽  
E Coli ◽  
Rapid Pcr ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Amino Acid Mutations ◽  
Bacterial Mutations

Background: Oral antibiotics, such as ciprofloxacin (CFX), are widely used for the treatment of acute and chronic pouchitis. Most bacterial mutations that confer quinolone resistance are at Ser-83 and Asp-87 in the gyrA gene and Ser-80 and Glu-84 in the parC gene. Methods: We obtained 51 stool samples from 43 patients who were diagnosed with ulcerative colitis and underwent ileal pouch-anal anastomosis. Patients were divided into 2 groups: 13 patients with CFX treatment of pouchitis and 30 patients without pouchitis. After extraction of fecal DNA, the amount of Escherichia coli 16S rRNA, gyrA, and parC gene DNA were measured using real-time polymerase chain reaction (PCR). Possible mutations at gyrA 83 and 87 and at parC 80 and 84 were investigated by PCR cloning and sequencing, and mutation rates were quantified by rapid PCR-restriction fragment length polymorphism. Results: Samples from both CFX-treated and -untreated patients had comparable levels of gyrA and parC gene DNA. Nucleic acid and amino acid mutations were identified at gyrA 83 and 87, and at parC 80 and 84. We successfully quantified mutation rates at gyrA 83 and 87, and at parC 84, all of which were significantly higher in samples from CFX-treated patients (70, 84, and 38%) than from CFX-untreated patients (13, 11, and 5%). Conclusion: E. coli in patient pouches may have mutations in their gyrA and parC genes that produce CFX resistance. Mutation rates of these genes were significantly higher in samples from CFX-treated patients. This study contributes to understanding the decrease and loss of CFX effectiveness against pouchitis.

scholarly journals Haemolytic uraemic syndrome and Shiga toxin-producing Escherichia coli infection in children in France

2000 ◽  
Vol 124 (2) ◽  
pp. 215-220 ◽  
Author(s):  
B. DECLUDT ◽  
P. BOUVET ◽  
P. MARIANI-KURKDJIAN ◽  
F. GRIMONT ◽  
P. A. D. GRIMONT ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Prospective Study ◽  
Shiga Toxin ◽  
Haemolytic Uraemic Syndrome ◽  
Serum Samples ◽  
E Coli ◽  
Escherichia Coli Infection ◽  
Stool Samples ◽  
Polymerase Chain

We conducted a study to determine the incidence of haemolytic uraemic syndrome (HUS) in children in France and to assess the role of Shiga-toxin-producing Escherichia coli (STEC) infection in the aetiology of HUS. In collaboration with the Société de Néphrologie Pédiatrique we undertook a retrospective review of all cases of HUS hospitalized from January 1993 to March 1995 and a 1-year prospective study (April 1995–March 1996) of epidemiological and microbiological features of cases of HUS. The polymerase chain reaction (PCR) procedure was used to detect stx, eae, e-hlyA genes directly from case stool samples. Serum samples from cases were examined for antibodies to lipopolysaccharide (LPS) of 26 major STEC serogroups. Two hundred and eighty-six cases were reported. The average incidence per year was 0·7/105 children < 15 years and 1·8/105 children < 5 years. During the prospective study, 122/130 cases were examined for evidence of STEC infection using PCR and/or serological assays and 105 (86%) had evidence of STEC infection. Serum antibodies to E. coli O157 LPS were detected in 79 (67%) cases tested. In conclusion, this study showed that STEC infection is an important cause of HUS in children in France, with a high proportion related to the O157 serogroup.

scholarly journals Prevalence, phylogeny, and antimicrobial resistance of Escherichia coli pathotypes isolated from children less than 5 years old with community acquired- diarrhea in Upper Egypt

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Rasha M. M. Khairy ◽  
Zahra Atef Fathy ◽  
Doaa Mohamed Mahrous ◽  
Ebtisam S. Mohamed ◽  
Soha S. Abdelrahim
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance Genes ◽  
E Coli ◽  
Extended Spectrum ◽  
Causative Agents ◽  
Typical Epec ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Lactamase Gene ◽  
Children Under 5

Abstract Background Diarrhoea, affecting children in developing countries, is mainly caused by diarrheagenic Escherichia coli (DEC). This study principally aimed to determine the prevalence of DEC pathotypes and Extended-spectrum β-lactamase (ESBL) genes isolated from children under 5 years old with diarrhea. Methods A total of 320 diarrhoea stool samples were investigated. E. coli isolates were investigated for genes specific for enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC), enteroinvasive E. coli (EIEC) and enterohemorrhagic E. coli (EHEC) using polymerase chain reaction (PCR). Furthermore, antimicrobial susceptibility testing, detection of antibiotic resistance-genes and phylogenetic typing were performed. Results Over all, DEC were isolated from 66/320 (20.6%) of the children with diarrhoea. EAEC was the predominant (47%), followed by typical EPEC (28.8%) and atypical EPEC (16.6%). Co-infection by EPEC and EAEC was detected in (7.6%) of isolates. However, ETEC, EIEC and EHEC were not detected. Phylogroup A (47%) and B2 (43.9%) were the predominant types. Multidrug-resistance (MDR) was found in 55% of DEC isolates. Extended-spectrum β-lactamase (ESBL) genes were detected in 24 isolates (24 blaTEM and 15 blaCTX-M-15). Only one isolate harbored AmpC β-lactamase gene (DHA gene). Conclusion The study concluded that, EAEC and EPEC are important causative agents of diarrhoea in children under 5 years. MDR among DEC has the potential to be a big concern.

Molecular detection of Escherichia coli (E. coli) from Diarrheal stool samples from children in Quetta, Balochistan, Pakistan.

2019 ◽  
Vol 2 (2) ◽  
pp. 27-31
Author(s):  
Tauseef M Asmat
Keyword(s):  
Escherichia Coli ◽  
Developing Countries ◽  
Disease Transmission ◽  
Causes Of Death ◽  
Growth Media ◽  
Huge Number ◽  
E Coli ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Control Disease

Diarrhea is one of the major causes of death in children, particularly in developing countries. Rapid detection and treatment is necessary to control disease transmission in the community and thus limiting the huge number of death toll. The major cause of diarrhea in developing countries is Escherichia coli (E. coli).This study was aimed to isolate E. coli from diarrheal stool samples from children aged 05 months to 05 years visited/ hospitalized in Quetta due to acute/persistent diarrhea. Diarrheal stool samples from 200 children were collected from Lady Sandeman Hospital Quetta and cultured on nutrient agar and later transferred to E. coli specific growth media for initial detection. For further confirmation the colonies were subjected to polymerase chain reaction (PCR). The PCR results revealed that 44(22%) samples out of 200 samples were positive for E. coli. These results indicate a high proportion of E. coli infection among children suffering with diarrhea.

scholarly journals Fecal carriage of extended-spectrum β-lactamase- and AmpC- producing Escherichia coli among healthcare workers

2015 ◽  
Vol 9 (03) ◽  
pp. 304-308 ◽  
Author(s):  
Rasha H. Bassyouni ◽  
Sylvana Nady Gaber ◽  
Ahmed Ashraf Wegdan
Keyword(s):  
Healthcare Workers ◽  
Control Measures ◽  
Carriage Rate ◽  
E Coli ◽  
Infection Control Measures ◽  
Fecal Carriage ◽  
Extended Spectrum ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Concentration Methods

Introduction: Commensal E. coli can be considered a reservoir of genes coding for antibiotic resistance that may be transmitted in hospitals by healthcare workers (HCWs). This study aimed to determine the fecal carriage rate of extended-spectrum β-lactamase (ESBL)-producing E. coli among HCWs. Methodology: Stool samples were collected from 200 HCWs. Phenotypic screening for ESBL and AmpC β-lactamases was performed using disk diffusion and minimum inhibitory concentration methods followed by the combined disks test and double synergy differential test for confirmation. Multiplex polymerase chain reaction (PCR) was used to detect blaSHV, blaTEM, blaCTX-M, and CIT groups for AmpC genes. Results: Of 200 E. coli isolates, 100% were susceptible to imipenem, and 59 (29.5%) were resistant to one or more third-generation cephalosporins. By molecular analysis, 21% (42/200) were colonized by ESBL-producing E. coli, and 3% (6/200) were colonized by AmpC-producing E. coli. The blaSHV gene was the predominant ESBL gene, detected in 81.8% of the resistant E. coli isolates. Conclusions: These findings highlight the increase in fecal carriage of E. coli carrying ESBL and AmpC genes among HCWs, which may be one of the causes of the spread of ESBL-producing bacteria in hospitals and requires sound infection control measures. This is the first study of the fecal carriage rate of E. coli carrying AmpC genes in HCWs.

scholarly journals High incidence of multidrug-resistant fecal E. coli producing ESBLs and carried ST131 in Jordanian adults

10.3823/0810 ◽  
2017 ◽  
Vol 7 (2) ◽  
Author(s):  
Reham Muin Abu Sneineh ◽  
Azmi Mahafzah ◽  
Nayef Abdallat ◽  
Asem A. Shehabi
Keyword(s):  
Multidrug Resistant ◽  
E Coli ◽  
Group I ◽  
High Incidence ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Tract Infections ◽  
The University ◽  
Hospital Acquired ◽  
St131 Clone

Background: Escherichia coli  is part of the human intestine normal flora, although it has the potential of causing variety of invasive and diarrheal diseases. It is also a frequent cause of community- and hospital-acquired urinary tract infections. Intestinal E. coli has the potential to develop rapidly multidrug resistant (MDR) and to emerge as extended-spectrum β-lactamases (ESBLs)-producer.    Methods: Over the period of July through November, 2015; 287 stool samples were collected from Jordanian adults who visited the students’ clinic of the University of Jordan. Fecal samples were collected and cultured for isolation of E. coli. The isolates were investigated for antimicrobial susceptibility, and molecular method of polymerase chain reaction (PCR) was performed for the detection genes of ST131 clone, blaCTX-M group I, blaCTX-M-15, blaNDM-1, blaVIM, blaIMP, blaOXA-48, blaKPC and fluoroquinolones resistance (gyrA and parC). Results: A total of 105/287 E. coli isolates (36.6%) were found to be multi-drug resistant (MDR) to at least 3 classes of antibiotics, of these 45.1% were ESBL-producers. A total of 51 representative MDR isolates indicated the following; 49% were found positive for ST131 clone, 58.8% were resistant for ciprofloxacin, and 41.2% were positive for CTX-M group I and CTX-M-15, respectively. All these MDR isolates were also positive for mutated both gyrA and ParC genes, and only 6 / 51 isolates (11.8%) were positive for each blaNDM-1 and blaKPC.  One out of 51 MDR isolates (2%) was positive for blaVIM, and none of these isolates was positive for blaIMP nor blaOXA-48 genes. Conclusion: This study indicated that a relatively high rates of commensal fecal E. coli isolates from Jordanian adults were MDR, ESBLs-producer and belonging to ST131 clone.  Also, high rates of CTX-M-15 and fluoroquinolones resistance were found among MDR E. coli isolates.    

scholarly journals Molecular detection of a new pathotype enteroaggregative haemorrhagic Escherichia coli (EAHEC) in Indonesia, 2015

2020 ◽  
Author(s):  
Wahyu Setyarini ◽  
Dadik Raharjo ◽  
Radita Yuniar Arizandy ◽  
Zakaria Pamoengkas ◽  
Subijanto Marto Sudarmo ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction Method ◽  
Unique Combination ◽  
Haemolytic Uremic Syndrome ◽  
E Coli ◽  
Virulence Potential ◽  
Uremic Syndrome ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Enterohemorrhagic E.Coli

Enteroaggregative haemorrhagic Escherichia coli (E. Coli, EAHEC) has been identified as the agent responsible for one of the largest outbreaks of gastroenteritis and Haemolytic-uremic syndrome (HUS) that is transmitted through food in Germany in 2011. The hypervirulent pathotype has a unique combination of two pathogens namely enterohemorrhagic E.coli strain (EHEC) which produces shiga/verotoxin and enteroaggregative E.coli toxins (EAEC) which produces toxins similar to ST and hemolysin. The toxin produced by the EAHEC strain is a hybrid pathotype that combines the virulence potential of the EAEC and EHEC strains that will damage the microcirculation, cause vasculitis and other toxic effects. The purpose of this study was to determine the percentage of samples infected with enteroaggregative hemorrhagic E. coli bacteria (EAHEC) in pediatric diarrhea patients at DR. Soetomo Hospital, Surabaya, Indonesia, 2015. This study used PCR (Polymerase Chain Reaction) method to detect enteroaggregative E. coli strains (CVD432 and aaic genes) and enterohemorrhagic E.coli (eae gene).The results showed that 33 out of 40 (82,5%) stool samples examined were detected enteroaggregative E. coli (EAEC), 4 out of 40 (10%) enterohemorrhagic E. coli (EHEC) and 3 out of 40 (7,5%) enteroaggregative haemorrhagic E. coli bacteria (EAHEC) , which caused diarrhea in pediatric diarrhea patients at Dr. Soetomo General Hospital. The unique combination of genomic features of the Surabaya outbreak strain, containing characteristics from pathotypes EAEC and EHEC, suggested that it represents a new pathotype enteroaggregative haemorrhagic E. coli (EAHEC). It is expected that development of specific primer design and sequencing are needed to continue in this research.

scholarly journals Molecular diagnosis of E.coliO157:H7 Which Isolated from Children with Diarrhea by using Multiplex PCR

2010 ◽  
Vol 7 (1) ◽  
pp. 207-215
Author(s):  
Baghdad Science Journal
Keyword(s):  
Escherichia Coli ◽  
Diagnostic System ◽  
Latex Agglutination ◽  
Diagnostic Methods ◽  
Bacterial Isolates ◽  
Biochemical Tests ◽  
E Coli ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Positive Results

A total of 96 stool samples were collected from children with bloody diarrhea from two hospitals in Baghdad. All samples were surveyed and examined for the presence of the Escherichia coli O157:H7 and differentiate it from other Non -Sorbitol Fermenting Escherichia coli (NSF E. coli). The Bacterial isolates were identifed by using morphological diagnostic methods, Samples were cultured on liquid enrichment medium, incubated at 37C? for 24 hrs, and then cultured on Cefixime Tellurite -Sorbitol MacConkey Agar (CT- SMAC). 32 non-sorbitol fermenting bacterial isolates were obtained of which 11 were identified as Escherichia coli by using traditional biochemical tests and API20E diagnostic system without differentiation between serotype O157:H7 and other NSF E. coli isolates . Four special biochemical tests were done for serotype O157:H7 differentiation from other NSF bacteria. Only 3 isolates belonging to the serotype O157:H7 were obtained . Latex agglutination test for O157 and H7 showed that the 3 isolates gave positive results with both tests. The Bacterial isolates were identifed by using Multiplex Polymerase Chain Reaction (MPCR) technology for the presence or absence of 4 genes (Stx1, Stx2, hlyA and eaeA) that encode for main virulence factors to diagnose E. coli O157:H7 isolated By using specific primers in MPCR . The result showed that one E. coli O157:H7 isolates contain all 4 genes , other isolates contain 3 genes: Stx2, hlyA & eaeA.

scholarly journals Detection of phenotypes, virulence genes and phylotypes of avian pathogenic and human diarrheagenic Escherichia coli in Egypt

2016 ◽  
Vol 10 (06) ◽  
pp. 584-591 ◽  
Author(s):  
Hazem Ramadan ◽  
Amal Awad ◽  
Ahmed Ateya
Keyword(s):  
Virulence Genes ◽  
Virulence Gene ◽  
Health Concern ◽  
E Coli ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Shiga Toxin Genes ◽  
Potential Public Health ◽  
Human Stool

Introduction: The purpose from this study was to determine phenotypes, intestinal virulence-associated genes, and phylotypic profiling of human diarrheagenic E. coli (DEC) and avian pathogenic E. coli (APEC). Methodology: A total of 108 chicken visceral organs (liver, spleen, heart) from 36 diseased birds (three organs per each bird) and 78 human stool samples (50 diarrheic patients and 28 healthy persons) were randomly collected during the first half of 2015 in the district of Mansoura city, Egypt. Conventional culturing, serotyping, and molecular characterization of virulence genes and phylogroups were performed. Results: Sixty-five (35%) biochemically identified E. coli isolates were detected from chicken visceral (29/108; 26.9%) and human stool samples (36/78; 46.2%). Serotypes O78, O2, and O1 were the most prevalent serotypes (62%) distinguished from APEC isolates, and only two similar serotypes (O119:H4 and O26:H11) were identified from both APEC and DEC isolates. By polymerase chain reaction (PCR), the respective percentages of 100 and 35 with eae and Shiga toxin genes were detected from APEC isolates while 50%, 27.8%, and 19.4% of human DEC isolates harbored eae, stx1, and stx2 genes, respectively. Phylogrouping revealed a significantly higher occurrence of pathogenic phylogroups (D and B2) in APEC (19/29; 65.5%) than in human DEC isolates (8/36; 22.2%). Conclusions: APEC isolates shared serotypes, virulence genes, and phylotypes with human DEC isolates, which is a subsequent potential public health concern. To the best of our knowledge, this is the first report in Egypt that determines virulence gene and phylogroup coexistence between APEC and DEC isolates.

scholarly journals High prevalence of qnrA and qnrB genes among fluoroquinolone-resistant Escherichia coli isolates from a tertiary hospital in Southern Nigeria

2021 ◽  
Vol 45 (1) ◽  
Author(s):  
Chibuzor M. Nsofor ◽  
Mirabeau Y. Tattfeng ◽  
Chijioke A. Nsofor
Keyword(s):  
Escherichia Coli ◽  
Susceptibility Testing ◽  
Tertiary Hospital ◽  
Vaginal Swab ◽  
Fluoroquinolone Resistance ◽  
E Coli ◽  
Diffusion Technique ◽  
High Prevalence ◽  
Polymerase Chain ◽  
And Control

Abstract Background This study was aimed to determine the prevalence of qnr genes among fluoroquinolone-resistant Escherichia coli (FREC) isolates from Nigeria. Antimicrobial susceptibility testing was performed by disc diffusion technique. Polymerase chain reaction was used to identify Escherichia coli (E. coli) and for the detection of qnr genes. Results A total of 206 non-duplicate E. coli were isolated from 300 clinical specimens analyzed. In all, 30 (14.6%) of these isolates were FREC; the resistance to fluoroquinolones among these 30 FREC showed 80% (24), 86.7% (26), 86.7% (26), 100% (30), 86.7% (26), 93.3% (28) and 86.7% (26) were resistant to pefloxacin, ciprofloxacin, sparfloxacin, levofloxacin, nalidixic acid, ofloxacin and moxifloxacin, respectively. The distribution of FREC among the various sample sources analyzed showed that 14%, 10%, 13.3%, 16.7% and 20% of the isolates came from urine, stool, high vaginal swab, endo cervical swab and wound swab specimens, respectively. More FREC were isolated from female samples 73.3% (22) compared to male samples 26.7% (8) and were more prevalent among the age group 26–35 years (40%). Twenty eight out of the 30 (93.3%) FREC isolates possessed at least one fluoroquinolone resistance gene in the form of qnrA 10 (33.3%) and qnrB 18 (60%), respectively; qnrS was not detected among the FREC isolates analyzed and 13.5% of the isolates possessed both the qnrA and qnrB genes. Phylogenetic analysis showed that these isolates were genetically diverse. Conclusions These findings suggest a possible resistance to fluoroquinolone is of high interest for better management of patients and control of antimicrobial resistance in Nigeria.

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