Simultaneous detection of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes using oscillatory-flow multiplex PCR
Simultaneous detection of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes using oscillatory-flow multiplex PCR
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使用振荡流多重 PCR 同时检测肠道沙门氏菌、大肠杆菌 O157:H7 和单核细胞增生李斯特氏菌
DOI：
10.1007/s00604-011-0584-5
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发表时间：
2011-06-01
影响因子：
5.7
通讯作者：
Xing, Da
中科院分区：
文献类型：
--
作者：
Wang, Haiying;Zhang, Chunsun;Xing, Da
摘要建立了毛细管微流控多重PCR检测多种食源性致病菌DNA的方法。PCR溶液以振荡方式通过三个温度区。研究了该微流控装置的热稳定性和样品蒸发性能。在控制条件下，一个高效的多重PCR完成了证明为278 bp，168 bp，和106 bp的DNA片段的同时扩增后35分钟内35个循环，同时检测沙门氏菌，大肠杆菌O 157：H7，李斯特菌。这比传统的PCR机器短得多。三种细菌基因组DNA的检测限分别为399、314和626拷贝/μL。这与用常规多重PCR获得的那些相当。因此，流动多重PCR技术具有良好的潜力，快速扩增和检测微生物食源性病原体的核酸。
图采用分流式多重PCR技术，可在35 min内同时扩增出278 bp、168 bp和106 bp的DNA片段，用于检测沙门氏菌。enterica、肠杆菌E. coliO 157：H7和L.单核细胞增多症这比传统的多重PCR短得多。
AbstractAn oscillatory-flow multiplex PCR method in a capillary microfluidic channel has been developed for the simultaneous determination of pre-purified DNA of multiple foodborne bacterial pathogens. The PCR solution passes three temperature zones in an oscillatory manner. The thermal stability and sample evaporation of the microfluidic device were investigated. Under controlled conditions, a highly efficient multiplex PCR was accomplished as demonstrated for the simultaneous amplifications of 278 bp, 168 bp, and 106 bp DNA fragments within 35 min after 35 cycles for simultaneous detection of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes. This is much shorter than that of a conventional PCR machine. The detection limits of bacterial genome DNA for the three species are about 399, 314, and 626 copies per μL, respectively. This is comparable to those obtained with the conventional multiplex PCR. Consequently, the oscillatory-flow multiplex PCR technology holds good potential for rapid amplification and detection of nucleic acids of microbial foodborne pathogens.
FigureBy using an oscillatory-flow multiplex PCR technology, the simultaneous amplifications of 278 bp, 168 bp, and 106 bp DNA fragments can be achieved in 35 min for detection of S. enterica, E. coli O157:H7, and L. monocytogenes. This is much shorter than that of a conventional multiplex PCR.