微流控免疫芯片富集捕获EHECO157︰H7的研究.docVIP

微流控免疫芯片富集/捕获EHEC O157︰H7 摘 要：目的　探讨基于免疫微珠的微流控芯片识别捕获EHEC O157︰H7EHEC O157︰H7μm的玻璃微珠上，再将微珠填充于PDMS/玻璃复合芯片μl/min和5min的最佳冲洗流速和冲洗时间下，该芯片在10min内可完成对EHEC O157︰H7EHEC O157︰H7 关键词：微流控芯片；免疫微珠；细菌捕获 中图法分类号：Q93-332；Q93-338 文献标识码：A A microfluidic chip to concentrating and capturing EHEC O157︰H7Department of Analytical Chemistry, College of Laboratory Medicine, Third Military Medical University, Chongqing 400038, China) Abstract: Objective To study the efficiency of concentration and capture of enterohemorrhagic Escherichia coli (EHEC) O157∶H7 by a self-made microfluidic chip based on microbead. Methods The anti-EHEC O157∶H7 antibody was first attached on the microbeads by direct covalent linkage, then the microbeads were filled into microfluidic chip. Thus, the bacteria capture chip was set up. Totally 1.6×109 CFU fluorescence labeled bacteria flowed into the chip at a sampling speed of 1, 2 and 5 μl/min respectively, and the elution speed of 0.5, 2, 5, 10 and 20 μl/min. The specific capture efficiency of the chip was strengthened through the optimization of sampling speed, elution speed and time. Results The best sampling speed, elution speed and time were 5 μl/min, 10 μl/min and 5 min respectively. There was significant difference between the test group and the control group (P0.01). Conclusion Our self-made bacteria capture chip is of high catching efficiency, which lay a good base for the construction of rapid detection microfluidic system of bacteria. Keywords: microfluidic chip; microbead; bacteria capture 肠出血性大肠埃希菌( Enterohemorrhagic Escherichia coli，EHEC) 是一种致病性大肠杆菌，O157∶H7 是EHEC 最主要的血清型，致病力强，最低感染剂量少于10 个病原菌，在世界各地多次发生暴发性感染，对人类健康形成巨大的威胁。所以，发展快速、敏感、特异的检测方法是预防和控制EHEC O157∶H7 传播和流行的重要手段[1,2]。在致病菌的检测中，用常规方法检测含量较低的样品时，为了达到检测方法所要求的检测限，往往需要对样品进行预富集处理。以细菌增殖为主的富集过程耗时过长，操作“十一五”课题(06MB248) Supported by the Research Project the “Eleventh Five-year Plan” for Medical Science Development of PLA(06MB248) 作者简介：管 潇，女，湖北省襄樊市人，硕士研究生，助理实验师，主要从事生物分析化学方面的研究。电话：(023)68752