Detection Of Toxigenic Clostridium Difficile In Stool By

粪便中产毒艰难梭菌的检测

• 批准号: 6675229
• 负责人: DANIEL P. FEDORKO
• 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/6675229
• 关键词: Clostridium difficile; DNA primers; antibiotics; antineoplastics; bacterial toxins; clinical research; diagnosis design /evaluation; diagnosis quality /standard; feces analysis; gastrointestinal disorder diagnosis; gastrointestinal drug absorption; genetic screening; hospital patient care; human tissue; method development; polymerase chain reaction; toxicant screening

Clostridium difficile-associated disease (CDAD) is a major problem for hospitalized patients recieving antibiotics or antineoplastic agents. Current labotatory methods for diagnosing CDAD include culture and assays for detection of the toxins produced by the organism or a cell-associated antigen. PCR assays for the diagnosis of CDAD have been reported in the liturature, but these reports are limited in scope and many of the primer pairs used are inefficient or amplify inappropriate portions of the C. difficile genome. We originally selected 5 primer pairs specific for the two toxin genes of C. difficile. We have selected the two primer pairs that are the most efficient in amplifying toxin A and toxin B. The base sequences for these two genes are both very AT rich, and we discovered that this prohibits the development of a sensitive Light Cycler assay. We have developed an ELISA format for our amplicon detection and are now in the process of determining the sensitivity of our assay before using patient specimens.

艰难梭菌相关性疾病（CDAD）是接受抗生素或抗生素治疗的住院患者的主要问题。目前用于诊断CDAD的实验室方法包括用于检测由生物体或细胞相关抗原产生的毒素的培养和测定。在文献中已经报道了用于诊断CDAD的PCR检测，但是这些报道在范围上是有限的，并且所使用的许多引物对是低效的或者扩增C. difficile基因组我们最初选择了5对针对C.很难我们已经选择了在扩增毒素A和毒素B中最有效的两个引物对。这两个基因的碱基序列都非常富含AT，我们发现这阻碍了灵敏的Light Cycler测定的发展。我们已经开发了一种用于扩增子检测的ELISA形式，目前正在使用患者标本前确定检测试剂盒的灵敏度。