scholarly journals Identification of Unconventional Intestinal Pathogenic Escherichia coli Isolates Expressing Intermediate Virulence Factor Profiles by Using a Novel Single-Step Multiplex PCR

2007 ◽  
Vol 73 (10) ◽  
pp. 3380-3390 ◽  
Author(s):  
Daniel Müller ◽  
Lilo Greune ◽  
Gerhard Heusipp ◽  
Helge Karch ◽  
Angelika Fruth ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Virulence Factor ◽  
Clinical Manifestations ◽  
Single Step ◽  
E Coli ◽  
Pathogenic Escherichia Coli ◽  
One Step ◽  
Differential Identification ◽  
Reference Strains

ABSTRACT Intestinal pathogenic Escherichia coli represents a global health problem for mammals, including humans. At present, diarrheagenic E. coli bacteria are grouped into seven major pathotypes that differ in their virulence factor profiles, severity of clinical manifestations, and prognosis. In this study, we developed and evaluated a one-step multiplex PCR (MPCR) for the straightforward differential identification of intestinal pathotypes of E. coli. The specificity of this novel MPCR was validated by using a subset of reference strains and further confirmed by PCR-independent pheno- and genotypic characterization. Moreover, we tested 246 clinical E. coli isolates derived from diarrhea patients from several distinct geographic regions. Interestingly, besides strains belonging to the defined and well-described pathotypes, we identified five unconventional strains expressing intermediate virulence factor profiles. These strains have been further characterized and appear to represent intermediate strains carrying genes and expressing factors associated with enteropathogenic E. coli, Shiga toxin-producing E. coli, enterotoxigenic E. coli, and enteroaggregative E. coli alike. These strains represent further examples of the extraordinary plasticity of the E. coli genome. Moreover, this implies that the important identification of specific pathotypes has to be based on a broad matrix of indicator genes. In addition, the presence of intermediate strains needs to be accounted for.

scholarly journals Molecular analysis of pathogenic Escherichia coli isolated from cow meat in Yogyakarta, Indonesia using 16S rRNA gene

2021 ◽  
Vol 22 (10) ◽  
Author(s):  
Alvita Indraswari ◽  
I Wayan Suardana ◽  
Aris Haryanto ◽  
Dyah Ayu Widiasih
Keyword(s):  
Escherichia Coli ◽  
Nucleotide Sequence ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Rrna Gene ◽  
Foodborne Disease ◽  
E Coli ◽  
Pathogenic Escherichia Coli ◽  
Cow Meat ◽  
Reference Strains

Abstract. Indraswari A, Suardana IW, Haryanto A, Widiasih DA. 2021. Molecular analysis of pathogenic Escherichia coli isolated from cow meat in Yogyakarta, Indonesia using 16S rRNA gene. Biodiversitas 22: 4566-4573. Meat has been recognized as a major source of foodborne disease and a public health problem. The characteristics of meat become an ideal growth medium for various microorganisms if not handled properly. Pathogenic Escherichia coli is one of the foodborne disease agents that causes diarrhea. Identification of pathogenic E. coli isolated from cow meat needs to be done. This research aims to study nucleotide sequence of 16S rRNA gene of pathogenic E. coli isolated from cow meat in Yogyakarta, Indonesia using Polymerase Chain Reaction (PCR). These fifteen isolates have been detected for eae target gene, then amplification of the 16S rRNA gene was carried out using primers 27F and 1492R. Phylogenetic tree reconstruction was performed on the fifteen isolates of pathogenic E. coli to figure out the relationship to reference strains available at the GenBank. Results show that nucleotide sequence among the fifteen isolates from different traditional markets in Yogyakarta, Indonesia and reference strains are very similar. The fifteen isolates have small genetic distance to the reference strains, and these fifteen isolates are also in the same clade with reference strains. This research shows that the fifteen isolates under investigation are closely related to the reference strains, which is Shiga-toxin producing E. coli (STEC). People should pay more attention in processing food stock, especially cow meat. Further research may focus on determining the strain of those fifteen isolates.

scholarly journals Molecular Detection and Identification of Intimin Alleles in Pathogenic Escherichia coli by Multiplex PCR

1999 ◽  
Vol 37 (8) ◽  
pp. 2719-2722 ◽  
Author(s):  
Sean D. Reid ◽  
David J. Betting ◽  
Thomas S. Whittam
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Molecular Detection ◽  
Pcr Assay ◽  
E Coli ◽  
Detection And Identification ◽  
Pathogenic Escherichia Coli

A multiplex PCR was designed to detect the eae gene and simultaneously identify specific alleles in pathogenicEscherichia coli. The method was tested on 87 strains representing the diarrheagenic E. coli clones. The results show that the PCR assay accurately detects eae and resolves alleles encoding the α, β, and γ intimin variants.

IDENTIFICATION OF THE AVIAN PATHOTYPE OF ESCHERICHIA COLI ON LAYER FLOCKS BY MULTIPLEX PCR

2019 ◽  
Vol 7 ◽  
pp. 905-911
Author(s):  
Marilena Burtan ◽  
Virgilia Popa ◽  
Maria Rodica Gurau ◽  
Doina Danes
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Virulence Genes ◽  
Virulence Gene ◽  
Economic Losses ◽  
Ompa Gene ◽  
Avian Pathogenic Escherichia Coli ◽  
E Coli ◽  
Pathogenic Escherichia Coli

Introduction: Colibacillosis in poultry is determined by avian pathogenic Escherichia coli (APEC) and represents an important source of economic losses  in the poultry industry. APEC’s pathogenicity relies on the presence and expression of different virulence factors. The genes ompA , iss  and  fimH, encoding the outer membrane protein, the protein inducing resistance to complement and the synthesis of type 1 fimbria are present in APEC strains. Objective: Escherichia coli strains isolated from layers were analysed to assess the pathotype they belong to. Methods: In order to detect the three genes associated with APEC strains, 16 E. coli isolates were investigated for virulence associated genes ompA, iss and fimH, using multiplex PCR. Results: From the 16 E.coli strains submitted, multiplex PCR assessment revealed that 14 (87.5%) of the E. coli strains isolated contained at least one virulence gene, while 2 (12.5%) strains did not harbour any of the virulence genes tested. The fimH gene was noted in 13 (81.25%) of the strains tested, the ompA gene has been present in 12 (75%) strains and the iss gene was present in 9 (56.25%) strains. Eight (50%) strains were found to present all three investigated genes. Conclusion: Presence of these genes is a strong indicatory to consider those strains as belonging to the APEC pathotype.

scholarly journals Effect of a lytic bacteriophage on rabbits experimentally infected with pathogenic Escherichia coli

2017 ◽  
Vol 25 (3) ◽  
pp. 273 ◽  
Author(s):  
J. Zhao ◽  
L. He ◽  
L. Pan ◽  
Y. Liu ◽  
H. Yao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Burst Size ◽  
Lethal Dose ◽  
Resistant Bacteria ◽  
Lytic Bacteriophage ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Pathogenic Escherichia Coli ◽  
One Step ◽  
The One

Pathogenic <em>Escherichia coli</em> (<em>E. coli</em>) is severely threatening the rabbit industry in China, and the concern over antibiotic-resistant bacteria has given rise to an urgent need for antibiotic alternatives. In this study, a member (ZRP1) of the <em>Myoviridae</em> family was isolated from rabbit faeces using a strain of rabbit atypical enteropathogenic <em>E. coli</em> (ZR1) as host. The one-step growth curve indicated that the latent period was around 25 to 30 min and the burst size was 144±31 plaque-forming unit/cell. The rate of phage-resistant mutation was 7×10<sup>–5</sup>±4×10<sup>–5</sup>. When the bacteriophage input at the multiplicity of infection (MOI) was 0.1, 1 or 10, the growth of host <em>E. coli</em> in broth was inhibited for 5 h. A single intravenous injection of ZRP1 at MOI 0.1, 1 or 10 significantly prolonged the survival time of rabbits which simultaneously received a lethal dose of ZR1.

scholarly journals OPTIMIZATION OF MOLECULAR DETECTION FOR Vibrio cholerae and PATHOGENIC Escherichia coli USING MULTIPLEX PCR

2021 ◽  
pp. e299
Author(s):  
Diana Elizabeth Waturangi ◽  
Jason Petrus ◽  
Rico Kosasih ◽  
Felicia Roseline
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Vibrio Cholerae ◽  
False Positive ◽  
Rapid Detection ◽  
Pathogenic Bacteria ◽  
Molecular Detection ◽  
False Negative ◽  
E Coli ◽  
Pathogenic Escherichia Coli

Vibrio cholerae and pathogenic Escherichia coli were considered as main causative agent foodborne diseases especially in many developing countries, such as Indonesia. Thereby, rapid detection of these pathogenic bacteria is necessary to treat food-borne related diseases causing by these bacteria. In this case, multiplex PCR allows multiple genes amplification in one reaction thereby enable to perform rapid detection of these pathogenic bacteria. The objective of this study is to optimize uniplex and multiples PCR of V. cholerae and pathogenic E. coli detection and determine the sensitivity and specificity of this assays. We used various virulence genes for each pathogenic bacterium as markers for uniplex and multiplex PCR detection. Based on this research, the optimum results of V. cholerae and pathogenic E. coli were obtained with a primer concentration of 16 µM for ctxA and ompU, 30 µM for ace, and 50 µM for zot, and toxR; 2 µM for elt and 5 µM for stx, respectively. Finally, based on the standardization method by ISO/TS 20836 these assays had 0% false positive, 0% false negative, 100% specificity, and 100% sensitivity; 0% false positive, 4% false negative, 100% specificity, and 96% sensitivity for V. cholerae and pathogenic E. coli respectively. The optimized method was qualified to be used as a molecular detection for V. cholerae as well as EHEC and ETEC detection according to ISO/TS 20836 (2017)  from drinking water samples.

Usefulness of a Multiplex PCR for Detection of Diarrheagenic Escherichia coli in a Diagnostic Microbiology Laboratory Setting

2016 ◽  
Vol 1 (2) ◽  
pp. 38-42 ◽  
Author(s):  
Khairun Nessa ◽  
Dilruba Ahmed ◽  
Johirul Islam ◽  
FM Lutful Kabir ◽  
M Anowar Hossain
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Clinical Laboratory ◽  
Proteinase K ◽  
Microbiology Laboratory ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay ◽  
Diagnostic Microbiology

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (<24 hrs) and less costly (<8.0 US $/ per test) but was less efficient in typing E. coli compared to proteinase K method (41.3% vs. 79.3% ; p<0.001). The sensitivity and specificity of boiling method compared to proteinase K method was 48.7% and 87.1% while the positive and negative predictive value was 93.5% and 30.7%, respectively. The majority of pathogenic E. coli were detected in children (78.0%) under five years age with 53.3% under one year, and 68.7% of the children were male. Children under 5 years age were frequently infected with EAEC (71.6%) compared to ETEC (24.3%), EPEC (2.7%) and STEC (1.4%). The multiplex PCR assay could be effectively used as a rapid diagnostic tool for characterization of diarrheagenic E. coli using a single reaction tube in the clinical laboratory setting.Bangladesh J Med Microbiol 2007; 01 (02): 38-42

Effect of Escherichia Coli Infection on Metabolism of Dietary Protein in Intestine

2020 ◽  
Vol 21 (8) ◽  
pp. 772-776
Author(s):  
Xiao-Pei Peng ◽  
Wei Ding ◽  
Jian-Min Ma ◽  
Jie Zhang ◽  
Jian Sun ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Intestinal Tract ◽  
Effector Proteins ◽  
Physical Injury ◽  
Dietary Proteins ◽  
Pathogenic Role ◽  
E Coli ◽  
Low Protein ◽  
Pathogenic Escherichia Coli ◽  
Escherichia Coli Infection

Dietary proteins are linked to the pathogenic Escherichia coli (E. coli) through the intestinal tract, which is the site where both dietary proteins are metabolized and pathogenic E. coli strains play a pathogenic role. Dietary proteins are degraded by enzymes in the intestine lumen and their metabolites are transferred into enterocytes to be further metabolized. Seven diarrheagenic E. coli pathotypes have been identified, and they damage the intestinal epithelium through physical injury and effector proteins, which lead to inhibit the digestibility and absorption of dietary proteins in the intestine tract. But the increased tryptophan (Trp) content in the feed, low-protein diet or milk fractions supplementation is effective in preventing and controlling infections by pathogenic E. coli in the intestine.

scholarly journals Avian Pathogenic Escherichia coli (APEC): An Overview of Virulence and Pathogenesis Factors, Zoonotic Potential, and Control Strategies

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 467
Author(s):  
Dipak Kathayat ◽  
Dhanashree Lokesh ◽  
Sochina Ranjit ◽  
Gireesh Rajashekara
Keyword(s):  
Escherichia Coli ◽  
Current Status ◽  
Neonatal Meningitis ◽  
Clear Understanding ◽  
Zoonotic Potential ◽  
Secretion Systems ◽  
Avian Pathogenic Escherichia Coli ◽  
E Coli ◽  
Pathogenic Escherichia Coli ◽  
Tract Infections

Avian pathogenic Escherichia coli (APEC) causes colibacillosis in avian species, and recent reports have suggested APEC as a potential foodborne zoonotic pathogen. Herein, we discuss the virulence and pathogenesis factors of APEC, review the zoonotic potential, provide the current status of antibiotic resistance and progress in vaccine development, and summarize the alternative control measures being investigated. In addition to the known virulence factors, several other factors including quorum sensing system, secretion systems, two-component systems, transcriptional regulators, and genes associated with metabolism also contribute to APEC pathogenesis. The clear understanding of these factors will help in developing new effective treatments. The APEC isolates (particularly belonging to ST95 and ST131 or O1, O2, and O18) have genetic similarities and commonalities in virulence genes with human uropathogenic E. coli (UPEC) and neonatal meningitis E. coli (NMEC) and abilities to cause urinary tract infections and meningitis in humans. Therefore, the zoonotic potential of APEC cannot be undervalued. APEC resistance to almost all classes of antibiotics, including carbapenems, has been already reported. There is a need for an effective APEC vaccine that can provide protection against diverse APEC serotypes. Alternative therapies, especially the virulence inhibitors, can provide a novel solution with less likelihood of developing resistance.

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.

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