期刊
ANALYTICA CHIMICA ACTA
卷 787, 期 -, 页码 93-101出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.aca.2013.05.043
关键词
Amino-modified silica-coated magnetic nanoparticles; Pathogens; Salmonella Enteritidis; Listeria monocytogenes; Polymerase chain reaction; Milk
资金
- Ministry of Science and Technology of China [2012AA101601, 2012BAK17B10, 2012BAD29B02]
- National Natural Science Foundation of China [31230058, 31000779, 31000063]
A method based on amino-modified silica-coated magnetic nanoparticles (ASMNPs) and polymerase chain reaction (PCR) was developed to rapidly and sensitively detect foodborne pathogens in raw milk. After optimizing parameters such as pH, temperature, and time, a trace amount of genomic DNA of pathogens could be extracted directly from complex matrices such as raw milk using ASMNPs. The magnetically separated complexes of genomic DNA and ASMNPs were directly subjected to single PCR (S-PCR) or multiplex PCR (M-PCR) to detect single or multiple pathogens from raw milk samples. Salmonella Enteritidis (Gram-negative) and Listeria monocytogenes (Gram-positive) were used as model organisms to artificially contaminate raw milk samples. After magnetic separation and S-PCR, the detection sensitivities were 8 CFU mL(-1) and 13 CFU mL(-1) respectively for these two types of pathogens. Furthermore, this method was successfully used to detect multiple pathogens (S. Enteritidis and L. monocytogenes) from artificially contaminated raw milk using M-PCR at sensitivities of 15 CFU mL(-1) and 25 CFU mL(-1), respectively. This method has great potential to rapidly and sensitively detect pathogens in raw milk or other complex food matrices. (C) 2013 Elsevier B.V. All rights reserved.
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