USE OF PCR RIBOTYPING FOR EPIDEMIOLOGICAL ANALYSIS OF CLOSTRIDIUM DIFFICILE

使用 PCR 核糖分型进行艰难梭菌流行病学分析

• 批准号: 3752278
• 负责人: C P CARTWRIGHT
• 金额: --
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3752278
• 关键词: Clostridium difficile; alleles; bacterial RNA; bacterial genetics; clinical chemistry; clostridial infection; diarrhea; epidemiology; genetic strain; microorganism classification; nosocomial infections; polymerase chain reaction; ribosomal RNA

Clostridium difficile is the principal cause of nosocomially acquired diarrhea in the U.S., and a number of large outbreaks of C. difficile- associated disease have been reported in the literature. To help track nosocomial acquisition and transmission in the Clinical Center, we have developed a simple, yet discriminatory technique for typing isolates of C. difficile. Conventional molecular approaches to typing microorganisms (for example plasmid analysis, restriction endonuclease analysis, and pulsed-field gel electrophoresis) are either relatively non-discriminatory, or time consuming, or both. To enable rapid assessment of the genetic relatedness of C. difficile isolates, we amplified the 16S to 235 intergenic spacer region of the ribosomal RNA gene cluster by PCR (PCR ribotyping). C. difficile possesses multiple alleles of this gene cluster, each differing in intergenic spacer length, and possession of the different alleles appears to vary in a 'strain' dependent manner. Use of this technique enabled us to assist the Hospital Epidemiology Service in identifying, retrospectively, an outbreak of C. difficile-diarrhea on 13W, and we are using the technique in a prospective manner to help identify future outbreaks.

艰难梭菌是医院获得性 腹泻在美国，以及一些大规模的C.艰难的，艰难的 相关疾病的报道在文献中。来帮助跟踪 在临床中心的医院获得和传播，我们 发明了一种简单但有区别性的打字技术 分离的C.很难传统的分子分型方法 微生物（例如质粒分析、限制性内切核酸酶分析）的方法 分析和脉冲场凝胶电泳）是相对 非歧视性的或耗时的，或两者兼有。以实现快速 对C.艰难分离株，我们 扩增核糖体16S至235基因间区， PCR核糖体分型（PCR ribotyping）。C. difficile拥有 该基因簇的多个等位基因，每个在基因间 间隔区长度和不同等位基因的拥有似乎有所不同 以依赖于“应变”的方式。这项技术的使用使我们能够 协助医院流行病学服务， 回顾性地，C.第13周出现腹泻， 正在以前瞻性的方式使用这项技术， 未来的爆发。