scholarly journals Detection of eae, bfpA, espA Genes on Diarrhoeagenic Strains of Escherichia coli Isolates

2015 ◽  
Vol 11 (1) ◽  
Author(s):  
Agnes Sri Harti ◽  
Susi Iravati ◽  
Widya Asmara
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Protein A ◽  
Dna Probes ◽  
Hybridization Technique ◽  
Secreted Protein ◽  
Enteropathogenic Escherichia Coli ◽  
E Coli ◽  
Typical Epec ◽  
Dna Fragment

The Enteropathogenic Escherichia coli (EPEC) is one of pathogenic strain of diarrheagenic E. coli group in children andinfant that occurs in developing countries. The significant virulence factors in pathogenic EPEC are eaeA (E. coli attachingeffacing), bfpA (bundle-forming pilus A) and espA (encoding secreted protein A) genes. The use of DNA probes to detect thevirulence genes in E. coli in Indonesia is not common yet. In this experiment the gene fragments of eae, bfpA, and espA were usedas probes to detect the EPEC among E. coli isolates from stool specimensin of diarrheic children attending Public Health Centersin Yogyakarta. The DNA samples were isolated from 49 diarrheagenic E. coli isolates. The DNA probes of eae, bfpA and espAwere obtained by amplification of DNA fragment of EPEC O126 using PCR technique. Furthermore, those probes were used toidentify the presence of those genes among E. coli isolates using hybridization technique. The results showed that 42 (85.7%)isolates were espA+, 25 isolates (51%) were eaeA+ (EPEC strains). Therefore among 25 isolates of EPEC, 20 isolates (80 %)among EPEC were bfpA+ (typical EPEC strains).Keywords : DNA probe, eae, bfpA, espA, EPEC.

Comparison of colony and stool blots for detection of enteropathogens by DNA probes

1991 ◽  
Vol 37 (5) ◽  
pp. 407-410
Author(s):  
Mônica A. M. Vieira ◽  
Beatriz E. C. Guth ◽  
Tânia A. T. Gomes
Keyword(s):  
Escherichia Coli ◽  
Key Words ◽  
Sensitivity And Specificity ◽  
Dna Probes ◽  
Type I ◽  
Enteropathogenic Escherichia Coli ◽  
Key Words Dna ◽  
E Coli ◽  
Heat Stable

DNA probes that identify genes coding for heat-labile type I (LT-I) and heat-stable type 1 (ST-I) enterotoxins, enteropathogenic Escherichia coli adherence factor (EAF), and Shigella-like, invasiveness (INV) are used to evaluate the sensitivity and specificity of stool blots in comparison with the sensitivity and specificity of colony blots in detecting enteropathoghens. The sensitivities of the probes in stool blots are 91.7% for the LT-I probe, 76.9% for the ST-I probes, 78.9% for the EAF probe, and 45.5% for the INV probe. The specificity of all probes is higher than 95%. In general, the stool blot method identifies as many if not more LT-I-, ST-I-, and EAF-producing E. coli infections than the colony blots. Key words: DNA probes, stool blots, enteropathogens, diagnosis.

scholarly journals Molecular and phenotypic typing of enteropathogenic Escherichia coli isolated in childhood acute diarrhea in Abuja, Nigeria

2017 ◽  
Vol 11 (07) ◽  
pp. 527-535 ◽  
Author(s):  
Casmir Ifeanyichukwu Cajetan Ifeanyi ◽  
Nkiruka Florence Ikeneche ◽  
Bassey Enya Bassey ◽  
Stefano Morabito ◽  
Caterina Graziani ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Acute Diarrhea ◽  
Antimicrobial Agents ◽  
Virulence Gene ◽  
Enteropathogenic Escherichia Coli ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Typical Epec ◽  
Marked Resistance

Introduction: Enteropathogenic Escherichia coli (EPEC) causes infectious diarrhea among children in developing countries. However, in Nigeria, due to limited laboratory resources, the genetic diversity of its virulence factors, which include intimin subtypes, remains undefined. Methodology: EPEC isolates from diarrheic children 60 months of age and younger in Abuja, Nigeria, were analyzed. Polymerase chain reaction (PCR) for EPEC virulence gene, Hep-2 cell adherence, and serotyping were performed. EPEC strains were further subtyped by PCR for the identification of intimin subtype genes α (alpha), β (beta), γ1 (gamma-1), and έ (epsilon). Antibiotic resistance and extended-spectrum beta-lactamase (ESBL) production was determined by Clinical and Laboratory Standards Institute guidelines. Results: Overall, 18 (4.5%) out of 400 children with acute diarrhea had EPEC infection.  Typical EPEC (tEPEC) strains were detected in 14 (3.5%), whereas 4 (1.1%) were atypical EPEC (aEPEC). A total of 15 (83.3%) of the EPEC isolated belonged to β intimin subtype gene, while the remaining 3 EPEC isolates possessed the intimin έ subtype. No α and γ intimin subtypes were detected. Traditional EPEC serotypes O114:H14 were detected only in tEPEC strains. Marked resistance to β-lactam agents were observed but no ESBL-producing tEPEC or aEPEC was detected. Conclusions: This is the first report of intimin subtype genes in Abuja, Nigeria. EPEC isolates of diverse serotypes resistant to β-lactam antimicrobial agents were observed. These data will be useful in facilitating the characterization of intimin variants of EPEC and some Shiga toxin-producing E. coli (STEC) in humans and other animal species.

scholarly journals Afa, a Diffuse Adherence Fibrillar Adhesin Associated with Enteropathogenic Escherichia coli

2002 ◽  
Vol 70 (5) ◽  
pp. 2681-2689 ◽  
Author(s):  
Rogéria Keller ◽  
Juana G. Ordoñez ◽  
Rosana R. de Oliveira ◽  
Luiz R. Trabulsi ◽  
Thomas J. Baldwin ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virulence Genes ◽  
Chromosomal Region ◽  
Enzyme Electrophoresis ◽  
Enteropathogenic Escherichia Coli ◽  
Epec Strain ◽  
Infantile Diarrhea ◽  
E Coli ◽  
Multilocus Enzyme Electrophoresis ◽  
Typical Epec

ABSTRACT O55 is one of the most frequent enteropathogenic Escherichia coli (EPEC) O serogroups implicated in infantile diarrhea in developing countries. Multilocus enzyme electrophoresis analysis showed that this serogroup includes two major electrophoretic types (ET), designated ET1 and ET5. ET1 corresponds to typical EPEC, whilst ET5 comprises strains with different combinations of virulence genes, including those for localized adherence (LA) and diffuse adherence (DA). Here we report that ET5 DA strains possess a DA adhesin, designated EPEC Afa. An 11.6-kb chromosomal region including the DA adhesin operon from one O55:H− ET5 EPEC strain was sequenced and found to encode a protein with 98% identity to AfaE-1, an adhesin associated with uropathogenic E. coli. Although described as an afimbrial adhesin, we show that both AfaE-1 and EPEC Afa possess fine fibrillar structures. This is the first characterization and demonstration of an Afa adhesin associated with EPEC.

scholarly journals Characterization of typical and atypical enteropathogenic Escherichia coli (EPEC) strains of the classical O55 serogroup by RAPD analysis

1999 ◽  
Vol 30 (4) ◽  
pp. 365-368 ◽  
Author(s):  
Dennys M. Girão ◽  
Sílvia Y. Bando ◽  
Valéria Brígido de C. Girão ◽  
Carlos A. Moreira-Filho ◽  
Sérgio Eduardo L. Fracalanzza ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Epidemiological Studies ◽  
Enteropathogenic Escherichia Coli ◽  
E Coli ◽  
Typical Epec ◽  
Rapd Method ◽  
Atypical Strains

The genetic diversity of 41 typical and atypical enteropathogenic Escherichia coli (EPEC) strains of the serogroup O55 was analyzed by using the random amplified polymorphic DNA (RAPD) method. All typical EPEC O55 strains were grouped in two clusters (A and C) and belonged to the serotype O55:H6, while cluster B included all atypical strains, which were of the serotype O55:H7. The three groups also included non-motile strains. RAPD may be a useful method for epidemiological studies on E. coli O55 infection.

scholarly journals Molecular detection of enteropathogenic Escherichia coli in asymptomatic captive psittacines

2012 ◽  
Vol 32 (9) ◽  
pp. 922-926 ◽  
Author(s):  
André Becker Saidenberg ◽  
Rodrigo Hidalgo F. Teixeira ◽  
Neiva Maria R. Guedes ◽  
Mariangela da Costa Allgayer ◽  
Priscilla Anne Melville ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Endangered Species ◽  
Disease Outbreaks ◽  
Ex Situ ◽  
Enteropathogenic Escherichia Coli ◽  
E Coli ◽  
Three Samples ◽  
In Captivity ◽  
Typical Epec ◽  
Conservation Of Endangered Species

Psittaciformes are one of the most endangered groups of birds, and several Brazilian species are classified between vulnerable and critically endangered. It is thus necessary to identify agents that cause infections in captive wild animals and to assess the risks posed thereof and to design interventions to minimize the possibility of disease outbreaks, leading to the conservation of endangered species. The purpose of this study was to identify enteropathogenic Escherichia coli (EPEC) cloacal isolates from asymptomatic psittacines in captivity and evaluate the distribution of the EPEC pathotype. Cloacal swabs were obtained from 46 asymptomatic birds, and resulting isolates were tested by polymerase chain reaction (PCR) for the presence of the attaching and effacing gene (eae) and bundle-forming pilus structural gene (bfpA) of EPEC. Samples from several species were tested, and three samples were found to be positive for the eae and bfpA genes and characterized as typical EPEC. This is the first report of this pathotype in asymptomatic psittacines. Although certain E. coli strains are more pathogenic than others, various factors should be considered when determining the potential of E. coli isolates to cause disease in captive psittacines. Birds that are positive for the EPEC (typical) strain could be zoonotic sources of infection, and may have acquired these strains through contact with humans or domestic animals. These findings may also be valuable for the long-term management of endangered species ex situ as one EPEC sample was isolated from a Red-tailed Amazon (Amazona brasiliensis).

scholarly journals Age-specific prevalence of diffusely adherent Escherichia coli in Brazilian children with acute diarrhoea

2008 ◽  
Vol 57 (3) ◽  
pp. 359-363 ◽  
Author(s):  
Liliana C. Spano ◽  
Ana Daniela I. Sadovsky ◽  
Paula N. Segui ◽  
Ketene W. Saick ◽  
Sonia M. S. Kitagawa ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Prospective Study ◽  
Blot Hybridization ◽  
Acute Diarrhoea ◽  
Dna Probes ◽  
E Coli ◽  
Typical Epec ◽  
Stool Specimens ◽  
A Prospective Study ◽  
Brazilian Children

In a prospective study between February 2003 and June 2004, stool specimens of children less than 2 years of age with diarrhoea (n=218) and without diarrhoea (n=86), living in Vitória, Espírito Santo, Brazil, were examined for the presence of diarrhoeagenic Escherichia coli. E. coli isolates were tested by colony blot hybridization with specific DNA probes designed to detect EPEC, ETEC, EIEC, EAEC, DAEC and EHEC/STEC. Diarrhoeagenic E. coli strains were detected as the sole pathogen in stools of 92 (30.3 %) children, including 72 (33.0 %) with diarrhoea and 20 (23.2 %) without diarrhoea. DAEC was the most frequent pathotype and was found significantly more often from patients (18.3 %) than from controls (8.1 %) (P<0.05), particularly among children more than 1 year of age (P=0.01). Atypical EPEC and EAEC isolates were isolated from both patients (5.5 % and 4.6 %, respectively) and controls (6.9 % and 6.9 %, respectively). ETEC was more frequently isolated from patients (3.2 %) than controls (1.2 %). Typical EPEC (0.9 %) and EIEC (0.4 %) isolates were detected only in children with diarrhoea. In conclusion, our data suggest that DAEC should be considered potential pathogens in the region of Brazil studied.

scholarly journals Molecular characterization and antimicrobial resistance of potentially human‐pathogenic Escherichia coli strains isolated from riding horses

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Pouya Reshadi ◽  
Fatemeh Heydari ◽  
Reza Ghanbarpour ◽  
Mahboube Bagheri ◽  
Maziar Jajarmi ◽  
...  
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Health Concern ◽  
Zoonotic Potential ◽  
Drug Resistant ◽  
Public Health Concern ◽  
E Coli ◽  
Pathogenic Escherichia Coli ◽  
Amoxicillin Clavulanic Acid

Abstract Background Transmission of antimicrobial resistant and virulent Escherichia coli (E. coli) from animal to human has been considered as a public health concern. This study aimed to determine the phylogenetic background and prevalence of diarrheagenic E. coli and antimicrobial resistance in healthy riding-horses in Iran. In this research, the genes related to six main pathotypes of E. coli were screened. Also, genotypic and phenotypic antimicrobial resistance against commonly used antibiotics were studied, then phylo-grouping was performed on all the isolates. Results Out of 65 analyzed isolates, 29.23 % (n = 19) were determined as STEC and 6.15 % (n = 4) as potential EPEC. The most prevalent antimicrobial resistance phenotypes were against amoxicillin/clavulanic acid (46.2 %) and ceftriaxone (38.5 %). blaTEM was the most detected resistance gene (98.4 %) among the isolates and 26.15 % of the E. coli isolates were determined as multi-drug resistant (MDR). Three phylo-types including B1 (76.92 %), A (13.85 %) and D (3.08 %) were detected among the isolates. Conclusions Due to the close interaction of horses and humans, these findings would place emphasis on the pathogenic and zoonotic potential of the equine strains and may help to design antimicrobial resistance stewardship programs to control the dissemination of virulent and multi-drug resistant E. coli strains in the community.

Spheroplasts of enteropathogenic Escherichia coli. I. Biological characteristics

1968 ◽  
Vol 14 (6) ◽  
pp. 675-678 ◽  
Author(s):  
B. Diena ◽  
R. Wallace ◽  
L. Greenberg
Keyword(s):  
Escherichia Coli ◽  
Tissue Culture ◽  
Biological Characteristics ◽  
Enteropathogenic Escherichia Coli ◽  
E Coli ◽  
O Antigen ◽  
Pathogenic Serotypes

The properties of glycine-induced spheroplasts of six pathogenic serotypes of E. coli were investigated. Fimbriae and flagella appeared to be only partially synthesized as was the somatic O antigen. Cytopathogenicity of these spheroplasts for tissue culture was reduced and the infection of the monolayers was retarded as compared with the normal bacillary forms. Sensitivity to phage was almost completely lost, suggesting that glycine had either interfered with the synthesis of phage receptors or had altered the mucopeptide layerwhich is the substrate for phage enzymes. Alternatively, the phage may become a prophage inside the spheroplast with the loss of virulence.

Expression and partial purification of human pro-opiomelanocortin in Escherichia coli

1989 ◽  
Vol 3 (2) ◽  
pp. 105-112 ◽  
Author(s):  
T. S. Grewal ◽  
P. J. Lowry ◽  
D. Savva
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Fusion Protein ◽  
Western Blot ◽  
Blot Analysis ◽  
Expression Vectors ◽  
Partial Purification ◽  
Amino Acid Residues ◽  
E Coli ◽  
Dna Fragment

ABSTRACT A large portion of the human pro-opiomelanocortin (POMC) peptide corresponding to amino acid residues 59–241 has been cloned and expressed in Escherichia coli. A 1·0 kb DNA fragment encoding this peptide was cloned into the expression vectors pUC8 and pUR291. Plasmid pJMBG51 (a pUC8 recombinant) was found to direct the expression of a 24 kDa peptide. The recombinant pUR291 (pJMBG52) was shown to produce a β-galactosidase fusion protein of 140 kDa. Western blot analysis showed that both the 24 kDa and 140 kDa peptides are recognized by antibodies raised against POMC-derived peptides. The β-galactosidase fusion protein has been partially purified from crude E. coli cell lysates using affinity chromatography on p-aminobenzyl-1-thio-β-d-galactopyranoside agarose.

scholarly journals The quantitative bacteriology of some commercial bivalve shellfish entering British markets

1975 ◽  
Vol 74 (3) ◽  
pp. 431-440 ◽  
Author(s):  
P. A. Ayres
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Health Risk ◽  
E Coli ◽  
Accepted Standard ◽  
Plate Counts ◽  
Bivalve Shellfish ◽  
Faecal Streptococci ◽  
Specific Illness

SUMMARYIncidents of non-specific illness associated with the consumption of oysters have highlighted the lack of published information on the bacteriology of shellfish suitable for consumption. Investigations showed that the majority of molluscan shellfish entering English markets conform to the accepted standard of less than 5 Escherichia coli/ml. tissue. The numbers of E. coli were related to the sanitary quality of the growing area but no relation could be established between numbers of E. coli and coliforms, faecal streptococci or Clostridium welchii. The numbers of non-specific bacteria varied considerably but shellfish from sources associated with non-specific illness yielded relatively high counts at 37° C. The results showed that there was no justification for a standard based on total plate counts, which often exceeded 106/g. Such a standard would have to be coupled with spoilage or the incidence of non-specific illness. The relation between the numbers of non-specific bacteria growing at 20 and 37° C. appears to be a useful measure for assessing the likelihood that raw shellfish are a public health risk.

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