Detection of Listeria monocytogenes based on combined aptamers magnetic capture and loop-mediated isothermal amplification. (March 2018)

• Record Type: Journal Article
• Authors: Feng, Junli; Dai, Zhiyuan; Tian, Xiaolan; Jiang, Xiaona
• Is Part Of: Food control. Volume 85(2018)
• Page Start: 443
• Page End: 452
• Publication Date: 2018-03
• Subjects: Listeria monocytogenes -- DNA aptamer -- Aptamers magnetic capture -- Loop-mediated isothermal amplification -- Rapid detection; Food -- Quality -- Periodicals; Food -- Analysis -- Periodicals; Food handling -- Periodicals; Food industry and trade -- Quality control -- Periodicals; Aliments -- Industrie et commerce -- Qualité -- Contrôle -- Périodiques; Aliments -- Qualité -- Périodiques; Aliments -- Analyse -- Périodiques; Hygiène alimentaire -- Périodiques; Food -- Analysis; Food handling; Food -- Quality; Periodicals; Electronic journals; 664.07
• DOI: 10.1016/j.foodcont.2017.10.027
• Languages: English
• ISSNs: 0956-7135

Abstract

Abstract: Rapid detection of Listeria monocytogenes ( L. monocytogenes ) is important to ensure timely treatment and to prevent outbreaks of listeriosis. Here, we reported an efficient and accurate system using aptamers magnetic capture and loop-mediated isothermal amplification (AMC-LAMP) for sensitive detection of L. monocytogenes . A set of combined aptamers (including four individual aptamers with high binding affinity to L. monocytogenes ) was conjugated to magnetic beads, used for capture L. monocytogenes . After incubation at room temperature for 45 min, the captured L. monocytogenes cells were directly used for DNA extraction (1 h). Subsequently, the LAMP assays were carried out at 63 °C for 40 min, and the amplification products were visualized via SYBR Green ® I staining. The AMC-LAMP had a detection limit of 5 CFU/mL, and the total assay time was approximate 3 h. By the protocols developed in this study, 17 food samples was test for L. monocytogenes without prior culture enrichment. The accuracy of AMC-LAMP was shown to be 100% when compared with culture-biotechnical examination, while PCR failed to detect L. monocytogenes in one of five positive samples. These results indicate the developed AMC-LAMP is a potential useful method for rapid screening and on-site detection of L. monocytogenes in food samples. Highlights: Combined aptamers were used to recognize and capture L. monocytogenes. Developed AMC-LAMP has a detection limit as low as 5 CFU/mL. The AMC-LAMP assay is suitable for rapid and on-site detection of L. monocytogenes.