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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50μm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 investigate the efficiency of EHEC O157: H7 capture by microfluidic chip based on immune microbeads. Methods Anti-EHEC O157:H7 antibody was immobilized on glass beads with a diameter of 50μm by chemical covalent attachment method. Then the beads were filled in PDMS / glass composite chips to establish bacteria controlled by fluid and immune reaction Sorting chip; through the optimization of the sample flow rate, flushing flow rate and flushing time, it can increase the chip-specific capture efficiency while reducing the nonspecific adsorption of the bacteria on the chip. Results The capture efficiency of EHEC O157:H7 was 40.00% -63.96% within 10min under the best flushing flow rate of 10μl / min and flushing time of 5min, and the experimental group and the control group were statistically Difference (P <0.01). Conclusion The EHEC O157: H7 identification chip constructed in this study has higher capture efficiency.