近年来，艰难梭菌感染（CDI）伴随着抗生素的滥用变得愈发盛行和严重，但是由于目前尚无一种敏感性、特异性较好，周转时间短、技术要求不高的诊断方法，故发展一种简易、快速、准确的CDI诊断技术具有重大的临床意义及现实需求。对临床的粪便样本采用核酸检测方法对艰难梭菌进行多重一体化检测是当前主要趋势，但是粪便样本处理过程复杂，核酸纯化效率不高，上游的核酸提取和下游的扩增检测的衔接，以及污染等问题是制约其在临床广泛应用的主要因素。本项目针对这些问题，拟通过建立粪便样本的快速处理流程，优化磁珠法粪便样本核酸提取和实时荧光多重PCR检测流程，使整个流程一体化和自动化；同时根据检测流程，设计新型的一体化封闭式检测卡盒，采用3D打印技术对其进行迭代设计和加工，使整个粪便样本的检测过程封闭在卡盒中完成，实现从样本输入到结果输出的C. diff多重一体化检测。最后通过对临床粪便样本的实际检测评估整个体系性能。

In recent years, Clostridium difficile infection (CDI) with the abuse of human antibiotics has become increasingly popular and serious, but there are no diagnostic methods with better sensitive, specificity, short turnaround time, low-demand technology at present, the development of a simple, fast and accurate CDI diagnosis new methods has significant clinical significance and practical needs. Nucleic acid testing for multiple and integrated detection of C. diff in clinical fecal samples is the main trend, but some factors like the complex process The development of a simple, fast and accurate new method for CDI diagnosis has significant clinical significance and practical needs. Nucleic acid testing for multiple and integrated detection of C. diff in clinical fecal samples is the main trend, but some factors like the complex process of the fecal samples, low nucleic acid purification efficiency, convergence of the upstream (nucleic acid extraction) and downstream (amplification and detection), and contamination problems restrict its application in clinical bioassays. In order to solve these problems, this project aims to establish rapid processing of fecal samples, optimize the process of nucleic acid extraction of fecal samples and real-time fluorescence multiplex PCR, and integrate and automate the whole process. At the same time, according to the integrated detection process, an integrated and closed cartridge is designed and processed by 3D printing technology iteratively. The whole detection process is completed in the cartridge to realize the C. diff multi-integration detection from the sample input to answer output. Finally, the performance of the entire system was assessed by the actual detection of clinical fecal samples.