CORE--LABORATORY

UTI核心实验室将提供必要的细胞培养， 年所有四个项目的微生物和分子分型程序 该计划。集中执行这些专门程序 确保它们的可用性、高质量和低成本。以下是 我们会提供以下服务： (1)分离和鉴定细菌的微生物学方法 项目1和项目3中的大肠杆菌和乳酸菌。 (2)培养(A)原代移行上皮细胞，用于 细菌鞘糖脂合成调控的研究 项目2、3和4中描述的依从性和细胞因子产生；以及 (B)由初级膀胱肌组成的膀胱器官结构。 与移行上皮细胞共同培养形成管状结构 作为项目3中导管感染的体外模型。 (3)分子分型技术，包括核糖体分型技术的应用 以及检测限制片段长度多态的方法，以测试 阴道和尿路分离的大肠埃希菌之间的遗传相关性 从参加研究的患者连续收集 项目1。 (4)使用分子遗传技术，包括聚合酶链式反应 菌落印迹与特异性DNA探针的反应和杂交 年从有症状的尿路感染妇女中采集的致尿系致病性大肠杆菌 项目1关于先前已知的大肠杆菌尿毒力决定因素 以及在项目3中发现的新发现的毒力因子。 (5)保存现有的致尿路病原性大肠杆菌菌株 和乳杆菌分离株以及建立更多的乳杆菌种质 并建立了铜绿假单胞菌菌种资料库 支持项目1、2、3和4。

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.