Detection Of Toxigenic Clostridium difficile In Stool By

粪便中产毒艰难梭菌的检测

• 批准号: 7004778
• 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/7004778
• 关键词: Clostridium difficile; DNA primers; antibiotics; antineoplastics; bacterial toxins; diagnosis design /evaluation; diagnosis quality /standard; enzyme linked immunosorbent assay; feces analysis; gastrointestinal disorder diagnosis; gastrointestinal drug absorption; genetic screening; hospital patient care; human tissue; method development; polymerase chain reaction; toxicant screening

We are evaluating the performance of a PCR assay designed in our laboratory for the diagnosis of Clostridium difficile disease. Our assay uses a primer set to detect the toxin A gene and another set to detect the toxin B gene. Endpoint detection of PCR products is an ELISA format. Our PCR assay will be compared to the following methods for diagnosis of C. difficile disease: toxigenic culture, cytotoxin assay, an ELISA for detection of toxins A and B, and a commercial PCR assay that detects the presence of the genes for both toxins A and B. All assays except our PCR assay have been performed on 144 stool specimens. We discovered that a unique toxin A deficient strain of C. difficile was not detected by our PCR assay. We have redisigned our toxin A gene primers and detection probe to allow us to detect this strain and have revalidated our PCR assay for detection of the toxin A gene. Once our PCR assay has been performed on all 144 specimens a chart review will be performed for specimens with conflicting results. Our results will demonstrate if PCR is a useful tool for diagnosis of C. difficile gastrointestinal disease. Note: This project has been merged with another project(CL010325-03 DLM) entitled "Evaluation of a Commercial PCR assay for Diagnosis of C. difficile Colitis."

我们正在评估我们实验室设计的用于诊断艰难梭菌疾病的PCR检测试剂盒的性能。我们的检测方法使用一组引物检测毒素A基因，另一组引物检测毒素B基因。PCR产物的终点检测是ELISA形式。我们的PCR检测方法将与以下诊断C的方法进行比较。难治性疾病：致突变培养、细胞毒素测定、用于检测毒素A和B的ELISA以及检测毒素A和B两者的基因的存在的商业PCR测定。除我们的PCR检测外，已对144份粪便标本进行了所有检测。我们发现了一种独特的毒素A缺陷型C.我们的PCR检测没有检测到艰难梭菌。我们重新设计了毒素A基因引物和检测探针，使我们能够检测该菌株，并重新验证了我们的PCR检测毒素A基因。对所有144份标本进行PCR检测后，将对结果不一致的标本进行图表审查。我们的结果将证明PCR是否是诊断C.艰难的胃肠道疾病。注：该项目已与另一个项目（CL 010325 -03 DLM）合并，该项目标题为“用于诊断C.艰难性结肠炎"