期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 255, 期 -, 页码 1455-1461出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2017.08.154
关键词
Emetic Bacillus cereus; Asymmetric polymerase chain reaction; Gold nanoparticles; Propidium monoazide
资金
- Training Plan for the Young Scientist (Jinggang Star) of Jiangxi Province, China [20142BCB23004]
- Research Foundation for Young Scientists of State Key Laboratory of Food Science and Technology, Nanchang University, China [SKLF-QN-201504]
Bacillus cereus is a causative agent of an emetic food-borne disease. In this study, visual detection of viable emetic B. cereus was developed using a propidium monoazide (PMA)-asymmetric polymerase chain reaction (asPCR) and unmodified gold nanoparticles (AuNPs). To detect only the viable emetic B. cereus, PMA treatment was selected before DNA extraction to eliminate the false-positive results from dead bacteria. In the presence of viable target bacteria, the long genomic DNA fragments from the cereulide synthetase gene (cesB) were produced by asPCR, which could be effectively absorbed onto naked AuNPs via coordination between Au and the nitrogen atoms of the exposed bases. After adding NaCl solution, visual detection with the naked-eye or with UV-vis spectrophotometer was possible within a few minutes. Under the optimum conditions, the limit of detection (LOD) for viable emetic B. cereus reached as low as 9.2 x 10(1) CFU/mL in 0.01 M phosphate-buffered saline and 3.4 x 10(2) CFU/mL in milk, which was adequate to meet the maximum limit imposed by the Commission Regulation (EC) No 2073/2005 (500 CFU/mL). The proposed method also exhibited excellent discrimination against 10 common pathogenic bacteria in milk. The PMA-asPCR-AuNPs colorimetric assay offers a promising application in the detection of low concentrations of food-borne pathogens. (C) 2017 Elsevier B.V. All rights reserved.
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