Multiplexed Bead-Based Mesofluidic System for Detection of Food-Borne Pathogenic Bacteria
Multiplexed Bead-Based Mesofluidic System for Detection of Food-Borne Pathogenic Bacteria
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DOI：
10.1128/aem.00854-09
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发表时间：
2009-11-01
影响因子：
4.4
通讯作者：
Ye, Bang-Ce
中科院分区：
文献类型：
--
作者：
Jin, Sheng-Quan;Yin, Bin-Cheng;Ye, Bang-Ce
在本研究中，一个简单和快速的多重珠为基础的介观流体系统（BMS）的同时检测食源性致病菌，包括金黄色葡萄球菌，副溶血性弧菌，单核细胞增生李斯特菌，沙门氏菌，坂崎肠杆菌，志贺氏菌，大肠杆菌O 157：H7，空肠弯曲菌。该系统是基于利用异硫氰酸酯改性的微珠，直径为250 μ m，这是固定与特定的氨基修饰的寡核苷酸探针，并放置在聚二甲基硅氧烷微通道。将来自所研究的病原体的PCR产物泵入微通道中，与经阿托伐他汀修饰的珠粒杂交，并使用常规微阵列扫描仪检测杂交信号。在1 pM和10 nM的浓度下，可以分别检测到细菌病原体特征性的核酸短序列（21个碱基）和PCR产物。该检测方法灵敏度高，特异性强，检测时间短于30 min。该方法简单、快速，研究的细菌种类的定量限在500至6，000 CFU/ml的范围内。通过对牛奶、鸡蛋、肉类等食品样品中细菌污染情况的分析，探讨了食源性细菌鉴定的可行性。结果表明，BMS方法可用于不同食品中多种病原体的有效检测。
In the present study, a simple and rapid multiplexed bead-based mesofluidic system (BMS) was developed for simultaneous detection of food-borne pathogenic bacteria, including Staphylococcus aureus, Vibrio parahaemolyticus, Listeria monocytogenes, Salmonella, Enterobacter sakazakii, Shigella, Escherichia coli O157:H7, and Campylobacter jejuni. This system is based on utilization of isothiocyanate-modified microbeads that are 250 mu m in diameter, which were immobilized with specific amino-modified oligonucleotide probes and placed in polydimethylsiloxane microchannels. PCR products from the pathogens studied were pumped into microchannels to hybridize with the oligonucleotide-modified beads, and hybridization signals were detected using a conventional microarray scanner. The short sequences of nucleic acids (21 bases) and PCR products characteristic of bacterial pathogens could be detected at concentrations of 1 pM and 10 nM, respectively. The detection procedure could be performed in less than 30 min with high sensitivity and specificity. The assay was simple and fast, and the limits of quantification were in the range from 500 to 6,000 CFU/ml for the bacterial species studied. The feasibility of identification of food-borne bacteria was investigated with samples contaminated with bacteria, including milk, egg, and meat samples. The results demonstrated that the BMS method can be used for effective detection of multiple pathogens in different foodstuffs.