微流控免疫芯片富集/捕获肠出血性大肠杆菌O157∶H7的研究  

A microfluidic chip to concentrate and capture EHEC O157∶H7

作　　者：管潇[1] 张惠静[1] 毕颖楠[1] 张莉[1] 郝敦玲[1] 韦劲宇[2] 

机构地区：[1]第三军医大学医学检验系分析化学教研室,重庆400038 [2]第三军医大学学员旅3队,重庆400038

出　　处：《第三军医大学学报》2009年第10期887-890,共4页Journal of Third Military Medical University

基　　金：全军医学科研"十一五"面上项目(06MB248)~~

摘　　要：目的探讨基于免疫微珠的微流控芯片识别捕获EHEC O157：H7的效率。方法采用化学共价连接的方法将抗EHEC O157：H7抗体固定于粒径为50txm的玻璃微珠上，再将微珠填充于PDMS／玻璃复合芯片中，建立由流体控制、免疫反应构成的细菌分选芯片；通过对进样流速、冲洗流速及冲洗时间的优化，在降低芯片对细菌的非特异性吸附的同时，增加芯片的特异性捕获效率。结果在10μl／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：H7的捕获，捕获效率达到40．00％-63．96％，且实验组与对照组有统计学差异（P〈0．01）。结论本研究构建的EHEC O157：H7识别芯片具有较高的捕获效率。Objective To study the efficiency of concentration and capture of enterohemorrhagic Escherichia coli（ EHEC ） O157：H7 by a self-made microfluidic chip based on mierobead. Methods The antiEHEC 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 × 10^9CFU 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 （P 〈 0. 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.

关 键 词：微流控芯片 免疫微珠 细菌捕获 

分 类 号：R318.04[医药卫生—生物医学工程] R378.21[医药卫生—基础医学]