Journal
MICROCHEMICAL JOURNAL
Volume 183, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.microc.2022.108063
Keywords
Ampicillin; Magnetic beads; L; monocytogenes; qPCR; Detection
Categories
Funding
- National Key R&D Program of China
- Research Foundation from Academic and Tech-nical Leaders of Major Disciplines in Jiangxi Province, China
- Research Project of State Key Laboratory of Food Science and Technology, Nanchang University, China
- [2018YFC1602500]
- [20194BCJ22004]
- [SKLF-ZZB- 202133]
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Listeria monocytogenes is a serious food-borne pathogen that poses a threat to global public health. This study successfully developed an Amp-MBs-qPCR strategy for the rapid and specific detection of L. monocytogenes in milk samples. The strategy demonstrated high sensitivity and specificity, with a capture efficiency higher than 90% and a limit of detection of 101-102 CFU/mL.
Listeria monocytogenes (L. monocytogenes) is a typical food-borne pathogen which poses a serious threat to global public health. Achieving rapid and sensitive detection of pathogens in food is critical to solve food safety problems. In this work, ampicillin functionalized magnetic beads (Amp-MBs) were synthesized and used as a capture probe for detection of L. monocytogenes in milk sample combined with quantitative real-time polymerase chain reaction (qPCR) with high sensitivity and specificity. Amp, as a broad spectrum antibiotic was selected to identify bacteria of the Listeria spp. powerfully, and the hly as a specific gene of L. monocytogenes was selected to establish qPCR for accurate detection. At optimal conditions, the capture efficiency of Amp-MBs for L. mono-cytogenes (101-106 CFU/mL) and other bacteria of Listeria spp. (105 CFU/mL) was higher than 90 %. The limit of detection of Amp-MBs-qPCR strategy for L. monocytogenes was 101 and 102 CFU/mL in PBS and artificially contaminated milk within 2.5 h, respectively. And this strategy could detect L. monocytogenes from high con-centration Listeria spp. with ascendant specificity. These results demonstrated that the Amp-MBs-qPCR strategy was successfully established and applied to the specific detection of L. monocytogenes in milk samples.
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