标题：Adherence patterns of diarrheagenic Escherichia coli strains isolated from children with gastrointestinal diarrhea and matched controls in a Hep-2 cell adhesion assay
期刊名称：Journal of Advances in Environmental Health Research
电子版ISSN：2345-3990
出版年度：2016
卷号：4
期号：3
页码：142-145
DOI：10.22102/JAEHR.2019.158736.1115
语种：English
出版社：Kurdistan University of Medical Sciences
摘要：Escherichia coli is considered as the main cause of epidemic outbreaks and endemic cases of child diarrhea. In the present study, the capacity of a large collection of Escherichia coli (E. coli) strains and matched controls was investigated to determine their adherence pattern on Hep-2 cell lines. A total of 66 pure E. coli strains, which were previously isolated from children aged less than 12 years with diarrhea, were selected for adherence assay. Moreover, 2 E. coli strains, with known adherence patterns, obtained from the Persian Type Culture Collection were used as the control. Hep-2 cell monolayers (50 to 70% confluence) were grown on glass cover slips. Subsequently, 20 µl of bacterial cultures (2 × 106 cfu/ml bacteria) was added into each well, and the plates were incubated in a humid atmosphere with 5% CO2 for 3 hours. Then, culture medium was aspirated from the monolayers, which were washed 4 times with phosphate-buffered saline, fixed in 70% aqueous methanol for 5 minutes, stained with Giemsa stain, and examined using light microscopy. In total, 66 E. coli strains were evaluated for their adherence pattern on Hep-2 cell line. All the isolates adhered to the Hep-2 cells with the exception of 3. The other 63 showed aggregative adherence (AA) (34%), localized adherence (LA) (31%), or diffuse adherence (DA) (29%) patterns. The present investigation revealed that this assay is convenient and economical for identification of a large number of diarrheagenic E. coli isolates’ adherence to Hep-2 cell lines.
关键词：Escherichia coli; Adherence Pattern; Cell Line; Children
