The UTI Core Laboratory will provide necessary cell culture, microbiological and molecular typing procedures for all four projects in the Program. Centralized performance of these specialized procedures assures their availability, high quality, and reduced cost. The following services will be provided: (1) Microbiological procedures for the isolation and characterization of E. coli and Lactobacillus in Projects 1 and 3. (2) Culture of (a) primary transitional epithelial cells for use in studies of the regulation of glycosphingolipid synthesis, bacterial adherence, and cytokine production described in Projects 2, 3, and 4; and (b) bladder organ structures consisting of primary bladder muscle co- cultured with transitional epithelial cells to form tubularized structures to be used as an in vitro model for catheter infections in Project 3. (3) The application of molecular typing techniques, including ribotyping and methods of detecting restriction fragment length polymorphism, to test for genetic relatedness among vaginal and urinary E. coli isolates serially collected from patients enrolled in the studies described in Project 1. (4) Use of molecular genetic techniques, including polymerase chain reaction and hybridization of colony blots with specific DNA probes to test uropathogenic E. coli collected from women with symptomatic UTI in Project 1 for previously recognized urovirulence determinants of E. coli as well as newly identified virulence factors found in Project 3. (5) Maintenance of existing collections of uropathogenic E. coli strains and Lactobacillus isolates and establishment of additional collections of these species as well as establishment of a P. aeruginosa strains bank to support Projects 1, 2, 3, and 4.
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