Simultaneous Detection of Yersinia Enterocolitica and Listeria Monocytogenes in Foodstuffs by Capillary Electrophoresis and Microchip Capillary Electrophoresis Laser-Induced Fluorescence Detector

• Published in: Journal of AOAC International Vol. 101; no. 6; pp. 1833 - 1838
• Main Authors: Li, Yongru, Su, Hongwei, Lan, Yajia
• Format: Journal Article
• Language: English
• Published: United States 01.11.2018
• Subjects: DNA, Bacterial - analysis; Electrophoresis, Capillary - methods; Fluorescence; Food Microbiology - methods; Limit of Detection; Listeria monocytogenes - genetics; Listeria monocytogenes - isolation & purification; Organic Chemicals - chemistry; Polymerase Chain Reaction - methods; Sensitivity and Specificity; Yersinia enterocolitica - genetics; Yersinia enterocolitica - isolation & purification
• ISSN: 1060-3271; 1944-7922
• DOI: 10.5740/jaoacint.17-0507

Food safety is one of the most important public health problems in the world, and pathogenic bacterium is a major factor causing serious foodborne diseases. Two methods of duplex PCR combined with capillary electrophoresis laser-induced fluorescence detector (CE-LIF) and microchip capillary electrophoresis laser-induced fluorescence detector (MCE-LIF) have been developed for the simultaneous detection of and in various foods. The specific conservative sequences of these two bacteria were amplified. After labelled with nucleic acid dye SYBR Gold and SYBR Orange, the PCR products were analyzed by CE-LIF and MCE-LIF, respectively. Under the optimal conditions, the detection of PCR products of the target bacteria was achieved in less than 15 min by CE-LIF and within 6 min by MCE-LIF. The alignment analysis demonstrated that the PCR products had good agreement with the sequences published in GenBank. The CE-LIF method could detect 10 CFU/mL and , and the MCE-LIF method could detect 100 CFU/mL and . The intraday precisions of migration time and peak area of DNA markers and PCR products were in the range of 1.13 to 1.18% and 1.60 to 6.29%, respectively, for CE-LIF and 1.18 to 1.48% and 2.85 to 4.06%, respectively, for MCE-LIF. : The proposed methods could be applied to target bacterial detection infood samples rapidly, sensitively, and specifically. : Two new methods based on CE and MCE have been developed for the simultaneous detection of and in foodstuffs, and they can detect the bacteria directly without any enrichment because of their high sensitivity.