Microbiology Core

本申请中的临床微生物学和实验室核心（微核心）代表了先前独立的微生物学实验室和人体挑战核心的融合。Micro Core将支持Cohen博士在项目1中的研究和埃尔金斯博士在项目2中的研究的标本采集、运输、储存和检测，包括检测泌尿生殖器标本中的淋病、衣原体和滴虫，以及检测生殖器溃疡病原体，特别是检测来自软下疳溃疡的杜克雷嗜血杆菌。Micro Core实验室人员执行市售的、FDA批准的核酸扩增检测分析，并在没有商业产品存在时开发和验证内部检测分析。微型核心的一个关键作用是按照项目1的要求，为马拉维利隆圭的性病检测提供培训和质量控制及保证计划。此外，我们将追求一个独立的具体目标， 表征临床标本中的杜克雷1- 1. II类菌株，该菌株建立在埃尔金斯博士最近发现的明显的第二类软下疳分离株的基础上，该分离株在当前申请的项目2的进展报告中进行了详细描述。检查I类和II类H的分布。为此，我们将收集和分析来自不同地理区域人群（包括马拉维、马达加斯加、喀麦隆、刚果和中国的性病诊所）的生殖器溃疡临床分离株。我们将通过PCR扩增长度多态性和dltA，dsrA和ncaA基因座的DNA序列分析来表征与两个谱系相关的基因型特征，并通过SDS PAGE和Western Blot进行表面蛋白表达和抗原特性的表型特征，以及体外抗生素敏感性测试。为了托马斯博士的研究 在项目3中，核心将用表达pilE、pilC和recA突变组合的淋病奈瑟菌变体进行人体挑战实验。微核心的资源包括在医院实验室，凡尔纳S。Caviness综合临床研究中心和传染病科。

The Clinical Microbiology and Laboratory Core (Micro Core) in this application represents a fusion of the previously separate Microbiology Laboratory and Human Challenge Cores. The Micro Core will support specimen collection, transport, storage and testing for Dr. Cohen's studies in Project 1 and Dr. Elkins' studies in Project 2 including detection of gonorrhea, chlamydia and trichomonas in urogenital specimens and detection of genital ulcer pathogens with special emphasis on Haemophilus ducreyi from chancroid ulcers. Micro Core lab personnel perform commercially available, FDA-approved nucleic acid amplification detection assays and develop and validate in-house detection assays when no commercial product exists. A key role of the Micro Core will be to provide training and quality control and assurance plans for STD testing in Lilongwe, Malawi as required in Project 1. In addition, we will pursue an independent specific aim to characterize 1-1.ducreyi Class II strains in clinical specimens, founded on Dr. Elkins' recent discovery of an apparent second class of chancroid isolates detailed in the progress report for Project 2 in the current application. To examine the distributions of Class I and Class II H. ducreyi strains for this aim, we will collect and analyze clinical isolates from genital ulcers from geographically diverse populations including STD clinics in Malawi, Madagascar, Cameroon, Congo and China. We will characterize genotypic features associated with the two lineages by PCR amplification length polymorphisms and DNA sequence analyses of the dltA, dsrA and ncaA loci, and phenotypic traits by SDS PAGE and Western Blot for surface protein expression and antigenic characteristics, and in vitro antibiotic susceptibility testing. For Dr. Thomas' studies in Project 3, the Core will conduct human challenge experiments with Neisseria gonorrhoeae variants expressing combinations of pilE, pilC and recA mutations. The resources of the Micro Core include facilities in the UNC Department of Hospital Laboratories, the Verne S. Caviness General Clinical Research Center and in the Division of Infectious Diseases.