scholarly journals Detection of Plasmid-Mediated qnr Genes Among the Clinical Quinolone-Resistant Escherichia coli Strains Isolated in Tehran, Iran

2018 ◽  
Vol 12 (1) ◽  
pp. 248-253 ◽  
Author(s):  
Reza Ranjbar ◽  
Sajjad S. Tolon ◽  
Mehrdad Sami ◽  
Reza Golmohammadi

Background:Escherichia coliis one of the most important bacterial agents to cause urinary tract infections. Inappropriate and unnecessary administration of antibiotics has led to an increase in the appearance of multidrug-resistantE. coliisolates, limiting treatment options. The increase in a number of resistant strains of bacteria is a major concern of health authorities worldwide.Objective:The purpose of this study was to determine the presence of theqnrgenes amongE. coliisolated from UTIs of patients in Baqiyatallah hospital in Tehran province, Iran.Method:Clinical urine samples of patients with suspected urinary tract infection were collected by standard methods in sterile disposable containers. After analysis of urine, microscopic observations and culture analysis, the bacterial genome was extracted by boiling method. PCR for detection ofqnrgenes includingqnrA,qnrBandqnrSwas done by specific primers, then PCR products were run using gel electrophoresis and visualized by gel documentation system.Results:In the present study among the 95 isolates, 60 strains were resistant to nalidixic acid. PCR showed that 92 strains were positive forqnrS. TheqnrAandqnrBgenes were not found among the clinical isolates.Conclusion:Our finding indicates a high level of resistance against nalidixic acid amongE. coliisolates recovered from the patients with UTI. Also, the high frequency ofqnrSimposes the importance of survey of molecular and genetic analysis of mechanisms of quinolone resistance inE. colistrains.

2014 ◽  
Vol 82 (4) ◽  
pp. 1572-1578 ◽  
Author(s):  
Karen L. Nielsen ◽  
Pia Dynesen ◽  
Preben Larsen ◽  
Lotte Jakobsen ◽  
Paal S. Andersen ◽  
...  

ABSTRACTCathelicidin (LL-37) and human β-defensin 1 (hBD-1) are important components of the innate defense in the urinary tract. The aim of this study was to characterize whether these peptides are important for developing uncomplicatedEscherichia coliurinary tract infections (UTIs). This was investigated by comparing urinary peptide levels of UTI patients during and after infection to those of controls, as well as characterizing the fecal flora of participants with respect to susceptibility to LL-37 andin vivovirulence. Forty-seven UTI patients and 50 controls who had never had a UTI were included. Participants were otherwise healthy, premenopausal, adult women. LL-37 MIC levels were compared for fecalE. coliclones from patients and controls and were also compared based on phylotypes (A, B1, B2, and D).In vivovirulence was investigated in the murine UTI model by use of selected fecal isolates from patients and controls. On average, UTI patients had significantly more LL-37 in urine during infection than postinfection, and patient LL-37 levels postinfection were significantly lower than those of controls. hBD-1 showed similar urine levels for UTI patients and controls. FecalE. coliisolates from controls had higher LL-37 susceptibility than fecal and UTIE. coliisolates from UTI patients.In vivostudies showed a high level of virulence of fecalE. coliisolates from both patients and controls and showed no difference in virulence correlated with the LL-37 MIC level. The results indicate that the concentration of LL-37 in the urinary tract and low susceptibility to LL-37 may increase the likelihood of UTI in a complex interplay between host and pathogen attributes.


2007 ◽  
Vol 1 (03) ◽  
pp. 257-262 ◽  
Author(s):  
Samuel Kariuki ◽  
Gunturu Revathi ◽  
John Corkill ◽  
John Kiiru ◽  
Joyce Mwituria ◽  
...  

Background: Uropathogenic Escherichia coli are increasingly becoming resistant to flouroquinolones and to other commonly available antimicrobials. We sought to investigate the genetic basis for fluoroquinolone and extended spectrum beta-lactam (ESBL) resistance in 17 fluoroquinolone-resistant (MIC of levofloxacin and ciprofloxacin >32 μg/ml) E. coli isolated from patients with urinary tract infections (UTIs). Methods: We applied PCR and Pulsed Field Gel Electrophoresis (PFGE) to characterize resistance genes and to determine clonal relatedness of strains, respectively. Results: Twelve of the 17 E. coli were resistant to multiple drugs, including ampicillin, co-amoxyclav, cefotaxime, ceftriaxone, ceftazidime and gentamicin and nalidixic acid and produced plasmid-mediated CTX-M-15 type ESBLs and CMY-2 AmpC type enzymes. The other 5 E. coli that were non-ESBL-producing were multiply resistant to ampicillin, nitrofurantoin, cefoxitin, nalidixic acid. Resistance to fluoroquinolones resulted from a combination of the presence of qnrA, qnrB, ciprofloxacin acetylating enzyme designated aac(6’)-1b-cr, and mutations in the two amino acid substitutions; 83 Serine (TCG) to Leucine (TTG) and 87 Aspartic acid (GAC) to Asparagine (AAC). Conclusion: Antibiogram patterns and PFGE of E. coli showed that these were community acquired UTI caused by pockets of clonally-related and some discreet strain types. Plasmid-mediated CTX-M-15 beta-lactamases and CMY-2 AmpC enzymes and fluoroquinolone resistant E. coli are becoming increasingly prevalent in hospitals in Kenya, posing a major challenge in the management of UTIs.


2021 ◽  
Vol 8 (9) ◽  
pp. 396-407
Author(s):  
Sheriff Wakil ◽  
Mustafa Alhaji Isa ◽  
Adam Mustapa

Multidrug resistance among Escherichia coli causing urinary tract infections (UTIs) and diarrhea are major public health problem worldwide which cause difficulty in treating the infections caused by Escherichia coli due to the high resistances. The study is aimed to determine the phenotypic and molecular detection of multidrug resistant E. coli isolated from clinical samples of patients attending selected Hospitals in Damaturu, Yobe State-Nigeria. Methods: Two hundred (200) clinical samples were collected aseptically from patient diagnosed with (100 stool samples) and UTI’s (100 urine samples) using sterile universal container. The samples were processed using standard microbiological methods for identification of E. coli. Samples were cultured on MacConkey agar (stool) and Cystine lactose electrolyte deficient agar (urine). The resulting colonies of isolates were further subculture on Eosin methylene blue agar for confirmatory and followed by gram stain, biochemical identification at Microbiology laboratory unit of Yobe State Specialist and Yobe State Teaching Hospital respectively. The antimicrobial susceptibility patterns were determined using Kirby-Bauer disc diffusion techniques and the phenotypic expression of extended spectrum beta-lactamases (ESBLs) were determined using modified double disc synergy test (MDDST) and also the three (3) resistance genes (blaTEM, accC1 and qnrA) were detected using polymerase chain reaction. Results: One hundred and twenty-two (122) isolates were resistant to antibiotics. The highest level of resistance was against amoxicillin (90.2%) while the least resistance was against sparfloxacin (24.3%). Thirty-seven (37) E. coli isolates shows MDR; the highest MDR was (24.3%) while least MDR was (5.4%). The PCR amplification of resistant genes (blaTEM, accC1 and qnrA) were detected on E. coli that shows positive ESBL and the bands were separated using agarose gel electrophoresis. Conclusion: The findings of this study show augmentin, ciprofloxacin and sparfloxacin are the most effective antibiotics against E. coli isolated from patients attending the two hospitals in Damaturu; who are diagnose with UTI and diarrheic infection. The resistant genes include; blaTEM, accC1 and qnrA coding for beta-lactam, aminoglycoside and quinolones were present in E. coli isolated from patients attending selected Hospitals in Yobe State, Nigeria. Keywords: Multidrug resistant, Escherichia coli, extended spectrum beta lactamase, resistance-associated genes, urinary tract infections, diarrheic.


2020 ◽  
Vol 5 (4) ◽  
pp. 176
Author(s):  
Purity Z. Kubone ◽  
Koleka P. Mlisana ◽  
Usha Govinden ◽  
Akebe Luther King Abia ◽  
Sabiha Y. Essack

We investigated the phenotypic and genotypic antibiotic resistance, and clonality of uropathogenic Escherichia coli (UPEC) implicated in community-acquired urinary tract infections (CA-UTIs) in KwaZulu-Natal, South Africa. Mid-stream urine samples (n = 143) were cultured on selective media. Isolates were identified using the API 20E kit and their susceptibility to 17 antibiotics tested using the disk diffusion method. Extended-spectrum β-lactamases (ESBLs) were detected using ROSCO kits. Polymerase chain reaction (PCR) was used to detect uropathogenic E. coli (targeting the papC gene), and β-lactam (blaTEM/blaSHV-like and blaCTX-M) and fluoroquinolone (qnrA, qnrB, qnrS, gyrA, parC, aac(6’)-Ib-cr, and qepA) resistance genes. Clonality was ascertained using ERIC-PCR. The prevalence of UTIs of Gram-negative etiology among adults 18–60 years of age in the uMgungundlovu District was 19.6%. Twenty-six E. coli isolates were obtained from 28 positive UTI samples. All E. coli isolates were papC-positive. The highest resistance was to ampicillin (76.9%) and the lowest (7.7%) to amoxicillin/clavulanic acid and gentamycin. Four isolates were multidrug-resistant and three were ESBL-positive, all being CTX-M-positive but SHV-negative. The aac(6’)-Ib-cr and gyrA were the most detected fluoroquinolone resistance genes (75%). Isolates were clonally distinct, suggesting the spread of genetically diverse UPEC clones within the three communities. This study highlights the spread of genetically diverse antibiotic-resistant CA-UTI aetiologic agents, including multidrug-resistant ones, and suggests a revision of current treatment options for CA-UTIs in rural and urban settings.


2019 ◽  
Vol 152 (Supplement_1) ◽  
pp. S131-S131
Author(s):  
Mohamed Abdelmonem ◽  
Amira Husseiny Mohamed ◽  
Hesham Shafik ◽  
Mohamed Shehata ◽  
Gamal EL-Sherbiny ◽  
...  

Abstract Escherichia coli (E coli) is one of the most commonly found pathogens in hospitals. Infections such as gastroenteritis, cystitis, urinary tract infections (UTI), meningitis, septicemia, and peritonitis were previously treated with broad-spectrum antibiotics. However, the emergence of infectious diseases involving multidrug-resistant (MDR) bacterial pathogens is still a major threat to human health. This study aimed to investigate the SulI (sulfonamide), TetA (tetracycline), and TetB resistance genes in E coli isolated from urine specimens from hospitalized patients. In the present cross-sectional study, a total of 55 strains of E coli were isolated from urine cultures of patients who had UTIs in ElKasr ElEiny and ELShorta hospitals. Samples were analyzed for bacteriological, biochemical examination, and agar disc-diffusion to evaluate their antibiotic susceptibility patterns. Polymerase chain reaction (PCR) method also was used to detect SulI, TetA/B genes by specific primers. The results suggested that E coli isolates were resistant to all multiple drugs used. Ampicillin showed the highest resistance of all the isolates followed by sulfonamide and tetracycline at 70%, 62%, and 53%, respectively. The lowest resistance detected with levofloxacin was 12%; however, there is no difference in the resistance pattern of gentamycin and aztreonam. The genotypes’ amplification revealed a positive correlation between SulI (sulfonamide) and TetA/B (tetracycline) resistance encoding genes and was shown in all the tested isolates as 100%. In our study, we found a mutation for sulfonamide and tetracycline genes in E coli that was isolated from UTI patients. The mutation is responsible for a multidrug-resistant strain due to the overuse of antibiotics. However, the World Health Organization recommends the use of trimethoprim-sulfamethoxazole and ampicillin as the first choice for UTI treatment. Our study recommends regulating and limiting the use of those antibiotics in order to minimize the dissemination of multidrug resistance for E coli.


mBio ◽  
2019 ◽  
Vol 10 (5) ◽  
Author(s):  
Nguyen Thi Khanh Nhu ◽  
Minh-Duy Phan ◽  
Brian M. Forde ◽  
Ambika M. V. Murthy ◽  
Kate M. Peters ◽  
...  

ABSTRACT Uropathogenic Escherichia coli (UPEC) is the major cause of urinary tract infections. Nearly half of all UPEC strains secrete hemolysin, a cytotoxic pore-forming toxin. Here, we show that the prevalence of the hemolysin toxin gene (hlyA) is highly variable among the most common 83 E. coli sequence types (STs) represented on the EnteroBase genome database. To explore this diversity in the context of a defined monophyletic lineage, we contextualized sequence variation of the hlyCABD operon within the genealogy of the globally disseminated multidrug-resistant ST131 clone. We show that sequence changes in hlyCABD and its newly defined 1.616-kb-long leader sequence correspond to phylogenetic designation, and that ST131 strains with the strongest hemolytic activity belong to the most extensive multidrug-resistant sublineage (clade C2). To define the set of genes involved in hemolysin production, the clade C2 strain S65EC was completely sequenced and subjected to a genome-wide screen by combining saturated transposon mutagenesis and transposon-directed insertion site sequencing with the capacity to lyse red blood cells. Using this approach, and subsequent targeted mutagenesis and complementation, 13 genes were confirmed to be specifically required for production of active hemolysin. New hemolysin-controlling elements included discrete sets of genes involved in lipopolysaccharide (LPS) inner core biosynthesis (waaC, waaF, waaG, and rfaE) and cytoplasmic chaperone activity (dnaK and dnaJ), and we show these are required for hemolysin secretion. Overall, this work provides a unique description of hemolysin sequence diversity in a single clonal lineage and describes a complex multilevel system of regulatory control for this important toxin. IMPORTANCE Uropathogenic E. coli (UPEC) is the major cause of urinary tract infections and a frequent cause of sepsis. Nearly half of all UPEC strains produce the potent cytotoxin hemolysin, and its expression is associated with enhanced virulence. In this study, we explored hemolysin variation within the globally dominant UPEC ST131 clone, finding that strains from the ST131 sublineage with the greatest multidrug resistance also possess the strongest hemolytic activity. We also employed an innovative forward genetic screen to define the set of genes required for hemolysin production. Using this approach, and subsequent targeted mutagenesis and complementation, we identified new hemolysin-controlling elements involved in LPS inner core biosynthesis and cytoplasmic chaperone activity, and we show that mechanistically they are required for hemolysin secretion. These original discoveries substantially enhance our understanding of hemolysin regulation, secretion and function.


2021 ◽  
Vol 10 (7) ◽  
pp. 414-418
Author(s):  
Greeshma Hareendranath

BACKGROUND Escherichia coli is one of the most important causes of urinary tract infections (UTIs). Increased antibiotic resistance may limit the therapeutic options for the treatment of these infections. Fosfomycin trometamol is a phosphonic acid derivative, which acts primarily by interfering with bacterial peptidoglycan synthesis with broad spectrum of activity against agents causing urinary tract infection with good antibiofilm activity and limited reports of resistance and hence is increasingly called upon for the treatment of multi drug resistant (MDR) organisms causing UTI. There are limited studies from India regarding the efficacy of this drug; so, the study was conducted to determine the in vitro efficacy of fosfomycin against uropathogenic MDR E. coli. METHODS This was a prospective study done in the Department of Microbiology, Government T.D. Medical College, Alappuzha, over a period of 1 year from April 2018 to March 2019. A total of 150 MDR urine samples were processed by routine microbiological methods and after identification of E. coli urinary isolates, antibiotic susceptibility testing was performed and results were interpreted following the Clinical and Laboratory Standards Institute guidelines (CLSI). Fosfomycin sensitivity was tested by the Kirby-Bauer disc diffusion method. RESULTS Among the 150 MDR urinary E. coli isolates, 148 (98 %) were sensitive to fosfomycin in our study. The susceptibility rate of fosfomycin was clearly higher than other commonly used drugs for UTI. All extended-spectrum beta-lactamases (ESBL) producing E. coli were sensitive to this drug. The susceptibility for nitrofurantoin was fair, whereas for ampicillin, norfloxacin, cefotaxime and trimethoprim / sulphamethoxazole was found poor. Relatively better rates of resistance were observed for parenteral antibiotics. CONCLUSIONS With an enormous increase in the bacterial pathogens resistant to first-line antibiotics, there has been a revival in the use of fosfomycin. The convenience of a single dose regimen, a good activity proven invitro, and minimal propensity for development of resistance pathogens makes fosfomycin an attractive regimen for the treatment of uncomplicated community and hospital acquired UTIs. In this regard, with the existing limited options for treating MDR organisms, fosfomycin finds its utility acting as an effective and promising option in the treatment of UTIs due to MDR pathogens in the future.


2017 ◽  
Vol 5 (4) ◽  
pp. 100-105 ◽  
Author(s):  
Mohadese Amiri ◽  
Maziar Jajarmi ◽  
Reza Ghanbarpour

Background: Antibiotic resistance (AR) is an important challenge in prevention, treatment and control of infectious diseases and is a public health threat for human. Escherichia coli strains are the major causes of urinary tract infections (UTIs). Objective: This research aimed to determine prevalence of resistance to quinolone and fluoroquinolone antibiotics and screen qnr genes among E. coli isolates from UTIs. Materials and Methods: A total of 105 E. coli isolates were obtained from UTI cases in Bojnord city (northeast of Iran) and confirmed by biochemical tests. All strains were studied to determine their resistance to 3 antibiotics including ciprofloxacin, nalidixic acid, and levofloxacin via disk diffusion method. Moreover, the frequency of qnrA, qnrB and qnrS genes and phylogroups was studied by conventional Polymerase chain reaction (PCR). Results: In this study, prevalence of phenotypic AR to ciprofloxacin, nalidixic acid and levofloxacin was 47.6%, 44.8% and 38.1%, respectively. Three isolates were positive for qnrS and 1 isolate was positive for qnrA. Seven phylogenetic groups were also identified as follows: 18% A0, 6.7% A1, 7.6% B1, 1.9% B22, 23.8% B23, 15.3% D1 and 26.7% D2. Conclusion: Prevalence of qnr genes was very low; thus, other types of qnr and plasmid-mediated quinolone resistance genes were probably responsible for the resistance. Phenotypic AR to the 3 antibiotics was found in approximately half of E. coli strains. It is strongly recommended that antibiogram tests should be done before prescribing the ciprofloxacin, nalidixic acid and levofloxacin for UTIs.


Author(s):  
Saskia-Camille Flament-Simon ◽  
Marie-Hélène Nicolas-Chanoine ◽  
Vanesa García ◽  
Marion Duprilot ◽  
Noémie Mayer ◽  
...  

Escherichia coli is the main pathogen responsible for extraintestinal infections. A total of 196 clinical E. coli consecutively isolated during 2016 in Spain (100 from Lucus Augusti hospital in Lugo) and France (96 from Beaujon hospital in Clichy) were characterized. Phylogroups, clonotypes, sequence types (STs), O:H serotypes, virulence factor (VF)-encoding genes and antibiotic resistance were determined. Approximately 10% of the infections were caused by ST131 isolates in both hospitals and approximately 60% of these infections were caused by isolates belonging to only 10 STs (ST10, ST12, ST58, ST69, ST73, ST88, ST95, ST127, ST131, ST141). ST88 isolates were frequent especially in Spain while ST141 isolates significantly predominated in France. The 23 ST131 isolates displayed four clonotypes: CH40-30, CH40-41, CH40-22 and CH40-298. Only 13 (6.6%) isolates were carriers of ESBL enzymes. However, 37.2% of the isolates were multidrug-resistant (MDR). Approximately 40% of the MDR isolates belonged to only four of the dominant clones (B2-CH40-30-ST131, B2-CH40-41-ST131, C-CH4-39-ST88 and D-CH35-27-ST69). Among the remaining MDR isolates two isolates belonged to B2-CH14-64-ST1193 i.e the new global emergent MDR clone. To our knowledge, it is the first identification of this emergent clone in Spain. Moreover, a hybrid ExPEC/enteroaggregative isolate belonging to A-CH11-54-ST10 clone was identified.


Author(s):  
Somayeh Bakhtiari ◽  
Hassan Mahmoudi ◽  
Sara Khosravi Seftjani ◽  
Mohammad Ali Amirzargar ◽  
Sima Ghiasvand ◽  
...  

Background and Objectives: Escherichia coli is the most common causative agent of urinary tract infections (UTIs) in 90-80% of patients in all age groups. Phylogenetic groups of these bacteria are variable and the most known groups are A, B1, B2 and D. The present study aimed to evaluate the phylogenetic groups of E. coli samples obtained from UTIs and their relation with antibiotic resistance patterns of isolates. Materials and Methods: In this study 113 E. coli isolates were isolated from distinct patients with UTIs referred to Hamadan hospitals. After biochemical and molecular identification of the isolates, typing and phylogenetic grouping of E. coli strains were performed using multiplex PCR targeting chu, yjaA and TSPE4.C2 genes. The anti-microbial susceptibility of the isolates to amikacin, ampicillin, trimethoprim-sulfamethoxazole, amoxicillin/clavulanic acid, ciprofloxacin, cefotaxime, imipenem, aztreonam, gentamicin, meropenem, nitrofurantoin, nalidixic acid and cefazolin was determined using disk diffusion method. Results: Of 113 isolates, 50 (44.2%), 35 (31%), 23 (20.4%) and 5 (4.4%) of samples belonged to group B2, group D, group A and group B1 phylogenetic groups respectively. All isolates were susceptible to meropenem, imipenem (100%), followed by amikacin (99.1%). The highest resistance rates were observed against ampicillin (74.3%) and nalidixic acid (70.8%). Correlation between phylogenetic groups and antibiotic susceptibilities was significant only with co-amoxiclav (P = 0.006), which had the highest resistance in phylogenetic group A. Conclusion: Prevalence of different phylogroup and resistance associated with them in E. coli samples could be variable in each region. Therefore, investigating of these items in E. coli infections, could be more helpful in selecting the appropriate antibiotic treatment and epidemiological studies.


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