scholarly journals Molecular Epidemiologic Approaches to Urinary Tract Infection Gene Discovery in Uropathogenic Escherichia coli

2000 ◽  
Vol 68 (4) ◽  
pp. 2009-2015 ◽  
Author(s):  
Lixin Zhang ◽  
Betsy Foxman ◽  
Shannon D. Manning ◽  
Patricia Tallman ◽  
Carl F. Marrs

ABSTRACT Urinary tract infection (UTI) is one of the most frequently acquired bacterial infections. The vast majority of UTIs are caused by a large, genetically heterogeneous group of Escherichia coli. This genetic diversity has hampered identification of UTI-related genes. A three-step experimental strategy was used to identify genes potentially involved in E. coli UTI transmission or virulence: epidemiologic pairing of a UTI-specific strain with a fecal control, differential cloning to isolated UTI strain-specific DNA, and epidemiologic screening to identify sequences among isolated DNAs that are associated with UTI. The 37 DNA sequences initially isolated were physically located all over the tester strain genome. Only two hybridized to the total DNA of the sequencedE. coli K-12 strain; eight sequences were present significantly more frequently in UTI isolates than in fecal isolates. Three of the eight sequences matched to genes for multidrug efflux proteins, usher proteins, and pathogenicity island insertion sites, respectively. Using population characteristics to direct gene discovery and evaluation is a productive strategy applicable to any system.

2006 ◽  
Vol 74 (6) ◽  
pp. 3427-3436 ◽  
Author(s):  
Simon Léveillé ◽  
Mélissa Caza ◽  
James R. Johnson ◽  
Connie Clabots ◽  
Mourad Sabri ◽  
...  

ABSTRACT Virulence factors of pathogenic Escherichia coli belonging to a recently emerged and disseminated clonal group associated with urinary tract infection (UTI), provisionally designated clonal group A (CGA), have not been experimentally investigated. We used a mouse model of ascending UTI with CGA member strain UCB34 in order to identify genes of CGA that contribute to UTI. iha was identified to be expressed by strain UCB34 in the mouse kidney using selective capture of transcribed sequences. iha from strain UCB34 demonstrated a siderophore receptor phenotype when cloned in a catecholate siderophore receptor-negative E. coli K-12 strain, as shown by growth promotion experiments and uptake of 55Fe complexed to enterobactin or its linear 2, 3-dihydroxybenzoylserine (DHBS) siderophore derivatives. Siderophore-mediated growth promotion by Iha was TonB dependent. Growth and iron uptake were more marked with linear DHBS derivatives than with purified enterobactin. The reported phenotype of adherence to epithelial cells conferred by expressing iha from a multicopy cloning vector in a poorly adherent E. coli K-12 host strain was confirmed to be specific to iha, in comparison with other siderophore receptor genes. iha expression was regulated by the ferric uptake regulator Fur and by iron availability, as shown by real-time reverse transcriptase PCR. In a competitive infection experiment using the mouse UTI model, wild-type strain UCB34 significantly outcompeted an isogenic iha null mutant. Iha thus represents a Fur-regulated catecholate siderophore receptor that, uniquely, exhibits an adherence-enhancing phenotype and is the first described urovirulence factor identified in a CGA strain.


2003 ◽  
Vol 71 (6) ◽  
pp. 3088-3096 ◽  
Author(s):  
Peter Redford ◽  
Paula L. Roesch ◽  
Rodney A. Welch

ABSTRACT Extraintestinal Escherichia coli strains cause meningitis, sepsis, urinary tract infection, and other infections outside the bowel. We examined here extraintestinal E. coli strain CFT073 by differential fluorescence induction. Pools of CFT073 clones carrying a CFT073 genomic fragment library in a promoterless gfp vector were inoculated intraperitoneally into mice; bacteria were recovered by lavage 6 h later and then subjected to fluorescence-activated cell sorting. Eleven promoters were found to be active in the mouse but not in Luria-Bertani (LB) broth culture. Three are linked to genes for enterobactin, aerobactin, and yersiniabactin. Three others are linked to the metabolic genes metA, gltB, and sucA, and another was linked to iha, a possible adhesin. Three lie before open reading frames of unknown function. One promoter is associated with degS, an inner membrane protease. Mutants of the in vivo-induced loci were tested in competition with the wild type in mouse peritonitis. Of the mutants tested, only CFT073 degS was found to be attenuated in peritoneal and in urinary tract infection, with virulence restored by complementation. CFT073 degS shows growth similar to that of the wild type at 37°C but is impaired at 43°C or in 3% ethanol LB broth at 37°C. Compared to the wild type, the mutant shows similar serum survival, motility, hemolysis, erythrocyte agglutination, and tolerance to oxidative stress. It also has the same lipopolysaccharide appearance on a silver-stained gel. The basis for the virulence attenuation is unclear, but because DegS is needed for σE activity, our findings implicate σE and its regulon in E. coli extraintestinal pathogenesis.


2020 ◽  
Vol 17 (35) ◽  
pp. 621-627
Author(s):  
May Ali Hussien UMRAN ◽  
Sumaya Najim AL-KHATEEB

The bacterium Escherichia coli is one of the best free-living organisms studied in depth. It is a surprisingly diverse species, since some strains of E. coli live in the intestine of animals as harmless commensals, while other distinct genotypes, such as an enteropathogenic or enterohemorrhagic E. coli, for example, cause morbidity and death marked as human intestinal pathogens. The purpose of this study was to develop and validate a PCR assay for a known and suspected uropathogenic E. coli virulence factor (kpsMT) gene region to determine the distribution of the gene and its role in the development of clinical diseases of the urinary system. A total of 25 urine samples were collected from patients with urinary tract infection (UTI) at Azadi and Kirkuk hospitals in the city of Kirkuk, Iraq. Samples of both genders and different ages were collected from patients with suspected urinary tract infection according to the clinical manifestations and symptoms diagnosed by the examining physician. The samples were cultured and positive samples were subjected to the IMViC test to identify E. coli bacteria and subsequently identified using the Vitek 2 compact system. Among 25 samples, 24 (96%) showed positive results for bacterial cultural growth. Of these, 17 (68%) were identified as Escherichia coli. Of the total of 17 isolates, 14 from patients with mild urinary tract infection, and 3 from patients with Urosepsis. The kpsMT gene was present in 14 isolates (82.3%), including 11 (78.5%) isolates from patients with mild urinary tract infection, and 3 (100%) isolates from patients with Urosepsis. It was concluded that Escherichia coli is the most prevalent in urine tract infection samples. Due to the abundance of the kpsMT gene in uropathogenic Escherichia coli (UPEC), this gene plays an important role in developing UTI if it is not treated correctly and quickly; mild cases of UTI can turn into Urosepsis.


2020 ◽  
Author(s):  
Mohammad Hasan Namaei ◽  
Hengameh Hamzei ◽  
Marzie Moghanni ◽  
Azadeh Ebrahimzadeh

Abstract Background: Urinary Tract Infection (UTI) is the most common bacterial infection in the world. E. coli is the predominant Pathogen. This study evaluates the prevalence of ESBL in E. colis isolated from patients with urinary tract infections with phenotypic and genotypic methods.Methods: This descriptive-analytical study was done on 155 isolates of E. coli isolated from patients with urinary tract infection who had received the study consent. After accurate identification of E. coli strains. ESBL production for Escherichia coli isolates which are resistant to ceftriaxone or ceftazidime was evaluated by CDT method. TEM, SHV and CTX-M genes were identified by PCR.Results: The results showed that 30 strains from 155 strains of E. coli had ESBL. Strains of ESBL producer were more in males was lower in educated persons. 38.9% of ESBL producer had antibiotic use, 29.9% -producing Escherichia hospitalization and 31.6% uti history. The highest level of drug allergy in the ESBL was related to nitrofurantoin, and the highest resistance was related to cefazolin, co-trimoxazole. The CTX-M and the CTX-M15 gene were found in 92.7% and 57.1% of cases, respectively; also the SHV and TEM genes were not found in any of ESBL-producing Escherichia coli strains. Most therapeutic response in patients was related to cefexime, ciprofloxacin and nitrofurantoin 27.4%, 26% 21.9%, respectively.Conclusion: This study showed that the history of antibiotic use, hospitalization, uti related to increase of ESBL-producing in E. coli isolates., the CTMX-M gene is the most common gene in ESBL-producing E. coli strains.


2020 ◽  
Vol 75 (12) ◽  
pp. 3656-3664
Author(s):  
Rasmus Richelsen ◽  
Jesper Smit ◽  
Henrik Carl Schønheyder ◽  
Pavithra Laxsen Anru ◽  
Belen Gutiérrez-Gutiérrez ◽  
...  

Abstract Objectives To assess the impact of ESBL production on mortality and length of hospital stay (LOS) of community-onset infections due to Escherichia coli or Klebsiella pneumoniae. Methods A population-based cohort study including all adult patients hospitalized with a first-time community-onset E. coli or K. pneumoniae bacteraemia or urinary tract infection in the North Denmark Region between 2007 and 2017. For each bacterial agent, we computed 1 year Kaplan–Meier survival curves and cumulative incidence functions of LOS, and by use of Cox proportional hazard regression we computed HRs as estimates of 30 day and 1 year mortality rate ratios (MRRs) and LOS among patients with and without ESBL-producing infections. Results We included 24 518 cases (among 22350 unique patients), of whom 1018 (4.2%) were infected by an ESBL-producing bacterium. The 30 day cumulative mortality and adjusted MRR (aMRR) in patients with and without ESBL-producing isolates was as follows: E. coli bacteraemia (n = 3831), 15.8% versus 14.0%, aMRR = 1.01 (95% CI = 0.70–1.45); E. coli urinary tract infection (n = 17151), 9.5% versus 8.7%, aMRR = 0.97 (95% CI = 0.75–1.26); K. pneumoniae bacteraemia (n = 734), 0% versus 17.2%, aMRR = not applicable; and K. pneumoniae urinary tract infection (n = 2802), 13.8% versus 10.7%, aMRR = 1.13 (95% CI = 0.73–1.75). The 1 year aMRR remained roughly unchanged. ESBL-producing E. coli bacteraemia was associated with an increased LOS compared with non-ESBL production. Conclusions ESBL production was not associated with an increased short- or long-term mortality in community-onset infections due to E. coli or K. pneumoniae, yet ESBL-producing E. coli bacteraemia was associated with an increased LOS.


2019 ◽  
Vol 2019 ◽  
pp. 1-8
Author(s):  
Sunayana Raya ◽  
Ankit Belbase ◽  
Laxmi Dhakal ◽  
Krishna Govinda Prajapati ◽  
Reena Baidya ◽  
...  

Background. Diabetic patients are more susceptible to urinary tract infection compared to nondiabetic patients, Escherichia coli being the most common uropathogen causing UTI. Unreasonable and incorrect antibiotic prescription for UTI in these patients may induce the development of antibiotic-resistant urinary pathogens resulting in delayed recovery and longer hospitalization. In addition to these, biofilm forming capacity of the pathogen may worsen the problem. The main aim of this cross-sectional study (conducted from March to September 2015) is to detect the biofilm forming capacity of UTI causing micro-organisms and compare the antibiotic resistance pattern of Escherichia coli, the most common cause of UTI, which will help the physician in choosing the best antibiotic. Method. Total of 1,099 clean-catch mid stream urine (CCMSU) was processed by standard microbiological technique; 182 were from the diabetic group and 917 nondiabetic. Following identification, all isolates were subjected to antibiotic susceptibility testing using modified Kirby-Bauer disc diffusion method. In-vitro biofilm forming capacity of the isolates were detected by Microtitre plate method. The data were analyzed using SPSS software 16. Result. Urinary tract infection was found to be significantly higher in diabetic patients (42.9%) compared to nondiabetic patients (17.4%) with Escherichia coli as the most common uropathogen in both diabetic and nondiabetic groups. Similarly, UTI was more common in elderly population (29.5%). Imipenem, nitrofurantoin and amikacin were found to be the most effective drug for uropathogenic E. coli in both diabetic and nondiabetic patients, whereas amoxicillin, ciprofloxacin, and cotrimoxazole were least effective. Of the total bacterial isolates, 43.3% showed positive results for in-vitro biofilm production by the Microtitre plate method. A significantly higher resistance rate was observed among biofilm producing E. coli for quinolones, cotrimoxazole, and third generation cephalosporin ceftriaxone. Most of the biofilm producers (79.5%) were found to be MDR (p-value 0.015). Conclusion. Elderly populations with diabetes are at a higher risk of UTI. Higher biofilm production and resistance to in-use antimicrobial agents in this study render its inefficacy for empirical treatment and point out the importance of biofilm screening to ensure the effective management of infection.


2009 ◽  
Vol 77 (8) ◽  
pp. 3312-3319 ◽  
Author(s):  
Te I. Weng ◽  
Hsiao Yi Wu ◽  
Pei Ying Lin ◽  
Shing Hwa Liu

ABSTRACT Escherichia coli is the most common cause of urinary tract infection. Elevated blood and urine interleukin-6 (IL-6) levels have been shown in inflammatory urinary tract diseases. The role of IL-6 in mediating the urodynamic dysfunction in response to E. coli-induced urinary tract infection has not yet been fully elucidated. In this study, we investigated the role of IL-6 in the nitric oxide (NO)-triggered alteration of contractile responses in the urinary bladder under an E. coli-induced inflammatory condition. The electrical field stimulation (EFS)-evoked contractions of the isolated detrusor strips, and immunoblotting for detecting protein expression in the bladders was measured short term (1 h) or long term (6 or 24 h) after intraperitoneal injection of E. coli endotoxin (lipopolysaccharide [LPS]) or intravesical instillation of human pyelonephritogenic E. coli-J96 (O4:K6) strain or LPS into mice. IL-6 and NO productions were increased in the urinary bladders of mice 1 to 24 h after LPS or E. coli-J96 treatment. Inducible NO synthase (iNOS) expression and protein kinase C (PKC) activation and EFS-evoked detrusor contractions were increased in the bladders at 6 h after LPS or E. coli-J96 treatment, which could be reversed by anti-IL-6 antibody and iNOS inhibitor aminoguanidine. At 1 h after LPS administration, bladder NO generation, endothelial NOS expression, and EFS-evoked detrusor contractions were effectively increased, whereas anti-IL-6 antibody could not reverse these LPS-induced responses. These results indicate that IL-6 may play an important role in the iNOS/NO-triggered PKC-activated contractile response in urinary bladder during E. coli or LPS-induced inflammation.


2008 ◽  
Vol 77 (3) ◽  
pp. 1155-1164 ◽  
Author(s):  
Mourad Sabri ◽  
Sébastien Houle ◽  
Charles M. Dozois

ABSTRACT Roles of the ZnuACB and ZupT transporters were assessed in Escherichia coli K-12 and uropathogenic E. coli (UPEC) CFT073. K-12 and CFT073 Δznu ΔzupT mutants demonstrated decreased 65Zn2+ uptake and growth in minimal medium. CFT073Δznu demonstrated an intermediate decrease of 65Zn2+ uptake and growth in minimal medium, whereas the CFT073ΔzupT mutant grew as well as CFT073 and exhibited a less marked decrease in 65Zn2+ uptake. CFT073 mutants grew as well as the wild type in human urine. In competitive infections in CBA/J mice, the ΔzupT mutant demonstrated no disadvantage during urinary tract infection. In contrast, the UPEC Δznu and Δznu ΔzupT strains demonstrated significantly reduced numbers in the bladders (mean 4.4- and 30-fold reductions, respectively) and kidneys (mean 41- and 48-fold reductions, respectively). In addition, in single-strain infection experiments, the Δznu and Δznu ΔzupT mutants were reduced in the kidneys (P = 0.0012 and P < 0.0001, respectively). Complementation of the CFT073 Δznu ΔzupT mutant with the znuACB genes restored growth in Zn-deficient medium and bacterial numbers in the bladder and kidneys. The loss of the zinc transport systems decreased both motility and resistance to hydrogen peroxide, which could be restored by supplementation with zinc. Overall, the results indicate that Znu and ZupT are required for growth in zinc limited-conditions, that Znu is the predominant zinc transporter, and that the loss of Znu and ZupT has a cumulative effect on fitness during UTI, which may in part be due to reduced resistance to oxidative stress and motility.


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