Microbiology Core

本申请中的临床微生物学和实验室核心(Micro Core)代表了以前分开的微生物学实验室和人类挑战核心的融合。Micro Core将支持科恩博士在项目1中的研究和埃尔金斯博士在项目2中的研究的标本收集、运输、存储和测试，包括检测泌尿生殖道标本中的淋病、衣原体和滴虫，以及检测生殖器溃疡病原体，特别是下巴溃疡中的杜氏嗜血杆菌。Micro Core实验室人员执行商业化的、FDA批准的核酸扩增检测分析，并在没有商业产品存在的情况下开发和验证内部检测分析。Micro Core的关键作用将是按照项目1的要求，为马拉维利隆圭的性病检测提供培训、质量控制和保证计划。此外，我们将追求独立的具体目标 在临床标本中鉴定1-1.Dureyi II类菌株，这是基于Elkins博士最近发现的明显的第二类软下巴分离物，在当前应用的项目2的进度报告中详细说明了这一点。为此，我们将从马拉维、马达加斯加、喀麦隆、刚果和中国等不同地区的人群中收集和分析生殖器溃疡的临床分离株，以检查I类和II类H.ducreyi菌株的分布。我们将通过dltA、DSRA和NCAA基因座的PCR扩增长度多态性和DNA序列分析，通过SDS PAGE和Western Blot分析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试验，来表征与这两个谱系相关的基因特征。托马斯博士的研究 在项目3中，核心将对淋病奈瑟菌的变种进行人类挑战实验，这些变种表达Pal、PilC和recA突变的组合。Micro Core的资源包括北卡罗来纳大学医院实验室部门、Verne S.Cavness综合临床研究中心和传染病部门的设施。

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.