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阀控多通道微流控芯片高通量快速检测大肠埃希菌 O157： H7

ISSN号：1001-764X
期刊名称：《临床检验杂志》
时间：0
分类：R446.5[医药卫生—诊断学;医药卫生—临床医学]
作者机构：[1]南昌大学研究生院医学部,南昌330006, [2]香港大学深圳医院医疗事务部,广东深圳518053, [3]中国科学院深圳先进技术研究院,广东深圳518055
相关基金：国家自然科学基金（61106128）;深圳发展改革委员会项目[（2012）385];深圳市科技计划项目（201101004）.

作者：张望[1,2], 徐小平[2], 高菊逸[1,2], 杜晶辉[1,2], 张宝月[3], 陈艳[3]

关键词：微流控芯片, 大肠埃希菌O157, H7, 快速检测, 高通量, microfluidic chip, Escherichia coli O157 ：H7 , rapid detection, high throughput

中文摘要：

目的建立一种基于气阀控制的多通道微流控芯片系统，实现高通量、低成本、快速检测大肠埃希菌O157：H7。方法采用集成多条反应通道和控制气阀的微流控芯片，用气阀控制芯片通道内反应的顺序，在通道交叉处构建出10个检测区。然后以免疫荧光技术为检测手段，检测水和粪便标本中的大肠埃希菌O157：H7结果成功建立了一种基于气阀控制的多通道微流控芯片系统，并应用于大肠埃希菌O157：H7的快速检测。此芯片一次可同时检测10个样本，单个样本检测时间少于10min、试剂消耗量仅为0．5斗L，对大肠埃希菌O157：H7的最低检测限为1×103CFU／mL。检测大肠埃希菌O157：H7组的荧光信号（4122±164）与大肠埃希菌ATCC25922（2．67±0．45）及伤寒沙门菌（3．33±0．94）比较，差异有统计学意义（t分别为35．78和30．79，P均〈0．01）。批内精密度为5．2％。将本法用于模拟临床样品中大肠埃希菌O157：H7的测定，敏感性和特异性分别为62％和100％，与培养法的总体符合率为81％。结论基于气阀控制的多通道微流控芯片系统可实现高通量、低成本、快速检测大肠埃希菌O157：H7，有望成为一种高效和快捷的新检测方法。

英文摘要：

Objective To establish a pneumatically controlled microfluidic muhichannel chip system for high throughput, low cost and rapid detection of Escherichia coli （E. coli） O157： H7. Method Utilizing the pneumatically controlled microfluidic chip with integrated multiehannels, the order of liquid flow of reaction on the chip was controlled by air valves, and ten detection areas were constructed at the crosses of channels. E. coli O157： H7 in water and stool specimens was then detected based on immunofluorescence. Results A pneumatically controlled multiehannel microfluidic chip was successfully constructed to detect E. coli O157： H7. Ten samples could be analyzed simultaneously in a single microfluidic chip. The average detection time for single sample was 10 minutes and the average rea- gent consumption was less than 0.5 μL. The minimum detectable limit for E. coli O157： H7 was 103 CFU/mL. The fluorescence signal of E. coli O157： H7（4 122 ± 164） was higher than those of E. coli ATCC 25922 （2.67±0.45） and Salmonella （3.33 ±0.94） with statis- tical significant difference （t = 35.78 and 30.79 respectively, P 〈 0.01 ）. Within-run precision was 5.2%. The detection for simula- ted clinical sample of E. coli O157 ： H7 by using the constructed chip showed sensitivity of 62% and specificity of 100%, and the total coincidence rate was 81% compared with the culture method. Conclusion The pneumatically muhichannel microfluidic chip based on micro air valves may achieve high-throughput, cost-effective and rapid detection for E. coli 0157 ： H7 and should be promising to become efficient, time-saving method for detection.

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