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A Novel High Throughput Assay For Measuring Bacterial Adhesion Activity to Epithelial Cells
Author(s) -
Zhao Qiuyan Julia,
Wei HuiLin,
Kondo Miwako,
Ou Boxin
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.lb255
Subject(s) - bioassay , adhesion , lysis , chemistry , potency , microbiology and biotechnology , chromatography , in vitro , biology , biochemistry , genetics , organic chemistry
Research studies show cranberry helps to prevent UTIs by inhibiting E. coli from adhering to the uroepithelium that line up the urinary tract. Currently, many cell based anti‐adhesion assays have been developed, however, they are time consuming and lacking reproducibility due to the inherent limitations of the assays. In this new assay, human uroepithelial cells T24 are plated in multi‐well plates, grown to confluence, rendered nonviable but adhered to plates by UVC treatment. Testing samples are added to the cells in TB media, followed by adding E. coli 1161. After co‐culturing, plates are washed, bacteria that remain to be adhered to the cells are lysed, and their ATP contents are read by a plate reader. The NIST SRM 3283 Cranberry Extract is a chosen reference standard, the anti‐adhesion index of a sample is obtained by fitting the sample's inhibition degree to the dose response curve of the NIST reference. The results are expressed as SRM 3283 equivalent per mg or per mL. Various phytochemicals from cranberry including their metabolites in biological fluids, plant extracts, and juices were tested by this new assay. Our results indicate that when compared to other fruits, cranberry demonstrated the highest potency of anti‐adhesion activity. The assay is reliable, high throughput, and can be utilized to test a wide of variety of samples.