食品中沙门菌和单增李斯特菌的多重PCR-芯片电泳快速检测

建立了食品中沙门菌和单增李斯特菌的多重PCR-芯片电泳快速检测方法。根据沙门菌和单增李斯特菌的特征基因合成2对特异性引物,优化聚合酶链反应(PCR)体系,采用芯片毛细管电泳快速检测食品中上述2种致病菌的多重PCR扩增产物。在优化的实验条件下,6 min内即可完成沙门菌和单增李斯特菌的同时检测;迁移时间的日内精密度为0.20%～1.7%,日间精密度3.7%～4.5%。

Rapid determination of salmonella and listeria monocytogenes in food samples by multiplex PCR-chip capillary electrophoresis

A rapid and simple method based on chip capillary system equipped with a laser-induced fluorescence detector was developed and applied to simultaneous analysis of the multiplex polymerase chain reaction(PCR) products of salmonella and Listeria monocytogenes in food samples.Two pairs of oligonucleotide primers were designed to exclusively amplify invA gene of Salmonella and hly gene of Listeria monocytogenes.The parameters of the multiplex PCR reaction and the chip capillary electrophoresis were optimized.The PCR products were separated and detected by the chip capillary system.The proposed method was employed to simultaneously detect the multiplex PCR products of two foodborne pathogenic bacteria under the optimal conditions within 6 min.The within-day relative standard deviations(RSD)of migration time was in the range of 0.20%～1.7% and the inter-day RSD were from 3.7%～4.5%.