Detection Of Toxigenic Clostridium Difficile In Stool By

艰难梭菌相关性疾病(CDAD)是接受抗生素或抗肿瘤药物治疗的住院患者的主要问题。目前诊断CDAD的实验室方法包括培养和检测由生物体或细胞相关抗原产生的毒素。用于诊断CDAD的聚合酶链式反应方法在文献中已有报道，但这些报告的范围有限，所使用的许多引物对效率低下或扩增艰难梭菌基因组的不适当部分。我们针对艰难梭菌的两种毒素基因选择了5对特异的引物。我们正在挑选在扩增靶标方面最有效的两对引物。然后，我们将开发一种使用Light Cycler的实时荧光聚合酶链式反应，并将使用该方法来评估利用粪便标本的PCR诊断CDAD的有效性。

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 have selected 5 primer pairs specific for the two toxin genes of C. difficile. We are in the process of selecting the two primer pairs that are the most efficient in amplifying their targets. We will then develop a realtime PCR using the Light Cycler and will use this assay to evaluate the utility of PCR for the diagnosis of CDAD using stool specimens.