建立基于正交的CRISPR的Clostridioides difficile毒素基因的新型检测平台
Tong Jiang1,2, Xinyi Hu1,2, Jilu Shen1,2
1The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China.
Microbiology spectrum
|June 28, 2023
概括
一个使用正交CRISPR技术的新OC-MAB遗传测试平台快速检测出Clostridioides difficile毒素基因. 这种方法为诊断C. difficile感染提供了高灵敏度和特异性,有助于感染控制.
科学领域:
- 分子生物学分子生物学
- 传染性疾病 传染性疾病
- 生物技术是生物技术.
背景情况:
- 困难菌是病房感染的主要原因,需要快速的诊断方法.
- 目前的诊断方法需要及时识别有效的治疗和感染控制.
研究的目的:
- 开发和评估OC-MAB平台,用于快速检测C. difficile毒素基因 (tcdA和tcdB) 的基因.
- 评估OC-MAB平台用于诊断C. difficile感染的敏感性,特异性和临床实用性.
主要方法:
- 开发OC-MAB (与多重重组合酶聚合酶放大相结合的直角CRISPR系统) 平台.
- 利用Cas13a和Cas12a酶进行特异性分离放大tcdA和tcdB基因产物.
- 采用双通道光检测通过qPCR和免疫染色体测试条用于视觉读取.
主要成果:
- OC-MAB平台表现出超高灵敏度,检测到tcdA和tcdB基因的水平低至10^1-10^2副本/mL.
- 在临床便样本中,基于光的方法与qPCR相比,实现了100%的灵敏度和特异性.
- 试验带方法显示了100%的灵敏度和96.3%的特异性,适合于点的护理测试.
结论:
- 正对角CRISPR技术为检测C. difficile毒素基因提供了强大而敏感的工具.
- OC-MAB平台提供了一种有前途的方法,可以快速,准确地诊断和控制C. difficile在医院获得的感染.
- 双目标侧流带格式非常适合于点的护理测试应用.
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