scholarly journals Detection of virulence genes of diarrheagenic Escherichia coli strains from drinking water in Khartoum State

2020 ◽  
Author(s):  
Ehssan H. Moglad ◽  
Omima Abdl El Jalil Adam ◽  
Maram M. Alnosh ◽  
Hisham N. Altayb
Keyword(s):  
Escherichia Coli ◽  
Drinking Water ◽  
Genomic Dna ◽  
Multiplex Polymerase Chain Reaction ◽  
Virulence Genes ◽  
Chain Reaction ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Polymerase Chain ◽  
Drinking Water Samples

Abstract This study aimed to determine the prevalence of virulence genes in all the diarrheagenic Escherichia coli DEC strains (EAEC, EHEC, EIEC, EPEC, and ETEC) isolated from drinking water from Khartoum State, Sudan. A total of 46 drinking water samples obtained from different water sources were analyzed for the presence of E. coli as fecal contamination indicator and the antimicrobial-resistant pattern of isolated E. coli DEC strain was investigated. The bacterial genomic DNA was used as a template for multiplex polymerase chain reaction (MPCR) for the detection of the EHEC (stx gene), EIEC (ipaH gene), EPEC (eae gene), and EAEC (aggR gene) as virulence and biomarker genes. Our results showed that ipaH gene was found in 41.3% (19/46) of isolates, and aggR gene detected in 30.4% (14/46) of isolates. Both aggR and ipaH were found positive in 9 (19.5%) isolates and as well the combination of aggR and stx genes were detected in 2 (4.3%) isolates. In conclusion, this report confirmed the presence of DEC strains in drinking water from different resources and locations. Such findings require separate future clinical research studies to examine waterborne pathogens that exist in this state's water and find a management solution to stop or avoid potential outbreaks.

scholarly journals High Frequency of Diarrheagenic Escherichia coli in HIV-Infected Patients and Patients with Thalassemia in Kerman, Iran

2015 ◽  
Vol 16 (4) ◽  
pp. 353-358 ◽  
Author(s):  
Hesam Alizade ◽  
Hamid Sharifi ◽  
Zahedeh Naderi ◽  
Reza Ghanbarpour ◽  
Mehdi Bamorovat ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Statistical Analysis ◽  
High Frequency ◽  
Multiplex Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Fecal Samples ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Polymerase Chain

This study was conducted on patients with thalassemia and HIV-infected patients to determine the frequency of diarrheagenic Escherichia coli in Kerman, Iran. We analyzed 68 and 49 E coli isolates isolated from healthy fecal samples of patients with thalassemia and HIV-infected patients, respectively. The E coli isolates were studied using a multiplex polymerase chain reaction to identify the enterotoxigenic E coli (ETEC), enterohemorrhagic E coli (EHEC), and enteropathogenic E coli (EPEC) groups. Statistical analysis was carried out to determine the correlation of diarrheagenic E coli between HIV-infected patients and patients with thalassemia using Stata 11.2 software. The frequency of having at least 1 diarrheagenic E coli was more common in patients with thalassemia (67.64%) than in HIV-infected patients (57.14%; P = .25), including ETEC (67.64% versus 57.14%), EHEC (33.82% versus 26.53%), and EPEC (19.11% versus 16.32%). The results of this study indicate that ETEC, EHEC, and EPEC pathotypes are widespread among diarrheagenic E coli isolates in patients with thalassemia and HIV-infected patients.

scholarly journals Diarrheagenic Escherichia coli pathotypes isolated from children with diarrhea in the Federal Capital Territory Abuja, Nigeria

2015 ◽  
Vol 9 (02) ◽  
pp. 165-174 ◽  
Author(s):  
Casmir Ifeanyichukwu Cajetan Ifeanyi ◽  
Nkiruka Florence Ikeneche ◽  
Bassey Enya Bassey ◽  
Nazek Al-Gallas ◽  
Ridha Ben Aissa ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virulence Genes ◽  
Diarrheal Disease ◽  
Cell Adherence ◽  
Chain Reaction ◽  
E Coli ◽  
Atypical Epec ◽  
Diarrheagenic Escherichia Coli ◽  
Stool Specimens ◽  
Polymerase Chain

Introduction: Escherichia coli are frequently isolated from diarrheic children in the Federal Capital Territory Abuja, Nigeria, but their virulent properties are not routinely evaluated. Therefore, the etiology of childhood diarrheal disease attributable to diarrheagenic Escherichia coli (DEC) in Abuja, Nigeria remains unknown. Methodology: Stool specimens from 400 acute diarrheic children between 0 and 60 months of age were studied.E. coli strains isolated were evaluated by polymerase chain reaction (PCR) for nine virulence genes and HEp-2 cell adherence to detect and identify five distinct diarrheagenic E. coli categories. Results: Diarrheagenic E.coli was detected in 51 (12.8%) of the diarrheic children. The observed DEC pathotypes were enteropathogenic E. coli (EPEC) in 18 (4.5%) children, enterotoxigenic E. coli (ETEC) in 16 (4.0%), enteroaggrative E. coli (EAEC) in 8 (2.0%), enterohaemorrhagic E. coli (EHEC) in 6 (1.5%), and enteroinvasive E. coli (EIEC) in 3 (0.8%). Four (1.0 %) EPEC strains with only the eae+ gene that adhered diffusely to HEp-2 cell were identified as atypical EPEC. All the DEC categories except atypical EPEC were identified in children between 6 and 12 months of age. Conclusions: This study underscores the need for routine evaluation of diarrheic children for virulence properties of infectious DEC. Atypical EPEC are emerging among the DEC pathotypes isolated from childhood acute gastroenteritis in Abuja, Nigeria.

Molecular Analysis of Uropathogenic E.coli Isolates from Urinary Tract Infections

2019 ◽  
Vol 19 (3) ◽  
pp. 322-326 ◽  
Author(s):  
Hassan Valadbeigi ◽  
Elham Esmaeeli ◽  
Sobhan Ghafourian ◽  
Abbas Maleki ◽  
Nourkhoda Sadeghifard
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Urinary Tract ◽  
Urinary Tract Infections ◽  
Multiplex Polymerase Chain Reaction ◽  
Virulence Genes ◽  
Uropathogenic Escherichia Coli ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Tract Infections

Introduction: The aim of the current study was to investigate the prevalence of virulence genes in uropathogenic Escherichia coli (UPEC) isolates in Ilam. Materials and Methods: For this purpose, a total of 80 UPEC isolates were collected for patients with UTIs during a 6 months period. The multiplex polymerase chain reaction (multiplex PCR) was used to detect the papEF, fimH, iucD, hlyA, fyuA, and ompT genes. Results: The prevalence of fimH, papEF, iucD, fyuA, hlyA, hlyA, and ompT genes were 87.5%, 47.5%, 60%, 67.5%, 27.5%, 47.5% and 71.2%, respectively. Among all of the isolates, 27 profiles were obtained. Conclusion: Our findings demonstrated that the most prevalence was found for fimH, and different distribution of virulence genes suggested different ability of pathogenicity.

scholarly journals Multiplex Polymerase Chain Reaction Assay for Identification of Enterotoxigenic Escherichia Coli Strains

2001 ◽  
Vol 13 (4) ◽  
pp. 308-311 ◽  
Author(s):  
Jacek Osek
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Multiplex Polymerase Chain Reaction ◽  
Direct Determination ◽  
Enteric Bacteria ◽  
Enterotoxigenic Escherichia Coli ◽  
Chain Reaction ◽  
Gram Negative ◽  
E Coli ◽  
Polymerase Chain

A multiplex polymerase chain reaction (PCR) system was developed for identification of enterotoxigenic Escherichia coli (ETEC) strains and to differentiate them from other gram negative enteric bacteria. This test simultaneously amplifies heat-labile (LTI) and heat-stable (STI and STII) toxin sequences and the E. coli-specific universal stress protein ( uspA). The specificity of the method was validated by single PCR tests performed with the reference E. coli and non- E. coli strains and with bacteria isolated from pig feces. The multiplex PCR allowed the rapid and specific identification of enterotoxin-positive E. coli and may be used as a method for direct determination of ETEC and to differentiate them from other E. coli and gram-negative enteric isolates.

scholarly journals Screening of AmpC-/ESBL-producing Escherichia coli isolates from livestock for STEC/EHEC virulence genes

2021 ◽  
Vol 72 (3) ◽  
pp. 3147
Author(s):  
F PEHLIVANOGLU
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Virulence Genes ◽  
Multidrug Resistant ◽  
Beta Lactamase ◽  
Chain Reaction ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Polymerase Chain ◽  
Flagellar Antigen

Livestock is an important reservoir of Shiga toxin-producing Escherichia coli and enterohemorrhagic E. coli (STEC/EHEC) strains and acts as a significant source of transmission to humans. In addition to the virulence of STEC/EHEC isolates, antibiotic resistance is also an escalating problem in these bacteria and increases the risk to public health. Therefore, the present study aimed to explore E. coli O157:H7 serotype and STEC/EHEC virulence genes in AmpC- and extended-spectrum beta-lactamase (ESBL)-producing E. coli isolates from cattle, chicken and sheep. A total of 61 confirmed AmpC- or ESBL-producing E. coli isolates were screened for the virulence genes (stx1, stx2, eae, ehxA, espP, katP and saa) and E. coli O157 (rfbO157) and H7 (fliCH7) genes by polymerase chain reaction (PCR). None of the ESBL-producing E. coli was positive for these genes, but six multidrug-resistant AmpC-producing E. coli were positive for the fliCH7 gene only. When considering the function of the H7 flagellar antigen of E. coli, it may be concluded that the development of ESBL/AmpC beta-lactamase production in the E. coli isolates with H7 flagella, which reside in the chicken intestine, may be potentially important for public health regarding both virulence and antimicrobial resistance.

scholarly journals PCR detection of Vibrio cholerae, Escherichia coli, and Salmonella sp. from bottled drinking water in Iran

2018 ◽  
Vol 12 (09) ◽  
pp. 700-705
Author(s):  
Mojtaba Bonyadian ◽  
Hamdallah Moshtaghi ◽  
Hanie Nadi
Keyword(s):  
Escherichia Coli ◽  
Drinking Water ◽  
Vibrio Cholerae ◽  
Screening Method ◽  
Chain Reaction ◽  
Culture Methods ◽  
E Coli ◽  
Drinking Waters ◽  
Polymerase Chain

Introduction: The quality of drinking water has an important role in human health. This study was aimed to detect Escherichia. coli, Salmonella sp. and Vibrio cholerae from bottled drinking waters produced in Iran. Methodology: A total of 240 samples of bottled water of different brands were collected for testing between March 2015 to December 2015 in Shahrekord-Iran. Samples were examined by polymerase chain reaction (PCR) combined with culture methods for the detection of E. coli, Salmonella sp., and V. cholerae. Results: The results of PCR revealed that the uidA gene from E. coli, IpaB gene from Salmonella sp, and epsM gene from V. cholerae were detected in 6 (2.5%), 1 (0.4 %), 0 (0%) of the samples, respectively. But in culture methods, only E. coli 5 (2.1%) were isolated from the samples. The contamination with E. coli was significantly higher (P < 0.05) in water produced during the hot seasons than the cold seasons. Conclusions: This study confirmed the presence of Escherichia coli as the main microorganism in bottle drinking water in Iran. Also, our study showed that PCR can be used as a screening method for monitoring the enteric pathogens in drinking water.

scholarly journals Detection of virulence genes and the phylogenetic groups of Escherichia coli isolated from dogs in Brazil

2018 ◽  
Vol 48 (2) ◽  
Author(s):  
Fernanda Morcatti Coura ◽  
Amanda Nadia Diniz ◽  
Carlos Augusto Oliveira Junior ◽  
Andrey Pereira Lage ◽  
Francisco Carlos Faria Lobato ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virulence Genes ◽  
Group Identification ◽  
Chain Reaction ◽  
Phylogenetic Groups ◽  
E Coli ◽  
Enterotoxin Genes ◽  
Virulence Factor Gene ◽  
Polymerase Chain ◽  
Cytotoxic Necrotizing Factor 1

ABSTRACT: This study identified the virulence genes, pathovars, and phylogenetic groups of Escherichia coli strains obtained from the feces of dogs with and without diarrhea. Virulence genes and phylogenetic group identification were studied using polymerase chain reaction. Thirty-seven E. coli isolates were positive for at least one virulence factor gene. Twenty-one (57.8%) of the positive isolates were isolated from diarrheal feces and sixteen (43.2%) were from the feces of non-diarrheic dogs. Enteropathogenic E. coli (EPEC) were the most frequently (62.2%) detected pathovar in dog feces and were mainly from phylogroup B1 and E. Necrotoxigenic E. coli were detected in 16.2% of the virulence-positive isolates and these contained the cytotoxic necrotizing factor 1 (cnf1) gene and were classified into phylogroups B2 and D. All E. coli strains were negative for the presence of enterotoxigenic E. coli (ETEC) enterotoxin genes, but four strains were positive for ETEC-related fimbriae 987P and F18. Two isolates were Shiga toxin-producing E. coli strains and contained the toxin genesStx2 or Stx2e, both from phylogroup B1. Our data showed that EPEC was the most frequent pathovar and B1 and E were the most common phylogroups detected in E. coli isolated from the feces of diarrheic and non-diarrheic dogs.

A Multiplex Polymerase Chain Reaction Assay for Rapid Detection and Identification of Escherichia coli O157:H7 in Foods and Bovine Feces†

2000 ◽  
Vol 63 (8) ◽  
pp. 1032-1037 ◽  
Author(s):  
PINA M. FRATAMICO ◽  
LORI K. BAGI ◽  
TIZIANA PEPE
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Multiplex Pcr ◽  
Multiplex Polymerase Chain Reaction ◽  
Ground Beef ◽  
Escherichia Coli O157 ◽  
Chain Reaction ◽  
E Coli ◽  
Polymerase Chain ◽  
Bovine Feces

A multiplex polymerase chain reaction (PCR) assay was designed to simplify detection of Escherichia coli O157:H7 and to identify the H serogroup and the type of Shiga toxin produced by this bacterium. Primers for a plasmid-encoded hemolysin gene (hly933), and chromosomal flagella (fliCh7; flagellar structural gene of H7 serogroup), Shiga toxins (stx1, stx2), and attaching and effacing (eaeA) genes were used in a multiplex PCR for coamplification of the corresponding DNA sequences from enterohemorrhagic E. coli (EHEC) O157:H7. Enrichment cultures of ground beef, blue cheese, mussels, alfalfa sprouts, and bovine feces, artificially inoculated with various levels of E. coli O157:H7 strain 933, were subjected to a simple DNA extraction step prior to the PCR, and the resulting amplification products were analyzed by agarose gel electrophoresis. Sensitivity of the assay was ≤1 CFU/g of food or bovine feces (initial inoculum level), and results could be obtained within 24 h. Similar detection levels were obtained with ground beef samples that underwent enrichment culturing immediately after inoculation and samples that were frozen or refrigerated prior to enrichment. The multiplex PCR facilitates detection of E. coli O157:H7 and can reduce the time required for confirmation of isolates by up to 3 to 4 days.

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