Use of DNA Probes and HEp-2 Cell Adherence Assay to Detect Diarrheagenic Escherichia coli

1988 ◽  
Vol 158 (1) ◽  
pp. 224-228 ◽  
Author(s):  
M. M. Levine ◽  
V. Prado ◽  
R. Robins-Browne ◽  
H. Lior ◽  
J. B. Kaper ◽  
...  
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 ◽  
...  

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.


2016 ◽  
Vol 1 (2) ◽  
pp. 38-42 ◽  
Author(s):  
Khairun Nessa ◽  
Dilruba Ahmed ◽  
Johirul Islam ◽  
FM Lutful Kabir ◽  
M Anowar Hossain

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


Author(s):  
Isaac Prah ◽  
Alafate Ayibieke ◽  
Samiratu Mahazu ◽  
Chihiro Tani Sassa ◽  
Takaya Hayashi ◽  
...  

2012 ◽  
Vol 79 (1) ◽  
pp. 411-414 ◽  
Author(s):  
Afonso G. Abreu ◽  
Vanessa Bueris ◽  
Tatiane M. Porangaba ◽  
Marcelo P. Sircili ◽  
Fernando Navarro-Garcia ◽  
...  

ABSTRACTAutotransporter (AT) protein-encoding genes of diarrheagenicEscherichia coli(DEC) pathotypes (cah,eatA,ehaABCDJ,espC,espI,espP,pet,pic,sat, andtibA) were detected in typical and atypical enteropathogenicE. coli(EPEC) in frequencies between 0.8% and 39.3%. Although these ATs have been described in particular DEC pathotypes, their presence in EPEC indicates that they should not be considered specific virulence markers.


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