Simultaneous and sensitive detection of three foodborne pathogens by multiplex PCR, capillary gel electrophoresis, and laser-induced fluorescence
Simultaneous and sensitive detection of three foodborne pathogens by multiplex PCR, capillary gel electrophoresis, and laser-induced fluorescence
复制标题
DOI：
10.1021/jf049038b
复制
发表时间：
2004-11-17
影响因子：
6.1
通讯作者：
Aznar, R
中科院分区：
文献类型：
--
作者：
Alarcón, B;García-Cañas, V;Aznar, R
同时检测金黄色葡萄球菌、单核细胞增生李斯特菌和沙门氏菌。已经接近了一个新的多重PCR为基础的程序，然后通过毛细管凝胶电泳与激光诱导荧光检测（多重PCR-CGE-LIF）。与平板凝胶电泳相比，使用CGE-LIF将多重PCR分析的灵敏度提高了10- 1000倍，允许检测2.6 x 10（3）cfu mL（-1）的S。金黄色葡萄球菌（L. aureus）570 cfu mL（-1）;在人工接种的食品中，未经富集的单核细胞增多症和沙门氏菌为790 cfu mL（-1）。富集6 h后，S.金黄色L.单核细胞增多症和沙门氏菌。CGE-LIF方法显示出重现性，提供的相对标准偏差（RSD）值低于0.8%的分析时间和低于5.8%的峰面积。多重PCR-CGE-LIF被证明是一种强大的分析工具，可以快速、重复性好、灵敏地同时检测各种食品病原体。
The simultaneous detection of Staphylococcus aureus, Listeria monocytogenes, and Salmonella spp. has been approached by a new multiplex PCR-based procedure followed by capillary gel electrophoresis with laser-induced fluorescence detection (multiplex-PCR-CGE-LIF). As compared to slab gel electrophoresis, the use of CGE-LIF improved from 10- to 1000-fold the sensitivity of the multiplex PCR analysis, allowing the detection of 2.6 x 10(3) cfu mL(-1) of S. aureus, 570 cfu mL(-1) of L. monocytogenes, and 790 cfu mL(-1) of Salmonella in artificially inoculated food, without enrichment. Following 6 h of enrichment, as low as 260, 79, and 57 cfu mL(-1) of S. aureus, L. monocytogenes, and Salmonella, respectively, were detected. The CGE-LIF method is shown to be reproducible, providing relative standard deviation (RSD) values lower than 0.8% for analysis time and lower than 5.8% for peak areas. The multiplex-PCR-CGE-LIF proved a powerful analytical tool to detect various food pathogens simultaneously in a fast, reproducible, and sensitive way.