A Rapid Method for Detection of Salmonella in Milk Based on Extraction of mRNA Using Magnetic Capture Probes and RT-qPCR
文献类型： 外文期刊
作者： Bai, Yalong 1 ; Cui, Yan 1 ; Suo, Yujuan 3 ; Shi, Chunlei 1 ; Wang, Dapeng 1 ; Shi, Xianming 1 ;
作者机构： 1.Shanghai Jiao Tong Univ, MOST USDA Joint Res Ctr Food Safety, Sch Agr & Biol, Shanghai, Peoples R China
2.Shanghai Jiao Tong Univ, State Key Lab Microbial Metab, Shanghai, Peoples R China
3.Shanghai Acad Agr Sci, Inst Agrifood Stand & Testing Technol, Shanghai, Peoples R China
关键词： amino-modified silica-coated magnetic nanoparticles; magnetic capture probes; Salmonella; RT-qPCR; milk
期刊名称：FRONTIERS IN MICROBIOLOGY （ 影响因子：5.64； 五年影响因子：6.32 ）
ISSN： 1664-302X
年卷期： 2019 年 10 卷
页码：
收录情况： SCI
摘要： Magnetic separation is an efficient method for target enrichment and elimination of inhibitors in the molecular detection systems for foodborne pathogens. In this study, we prepared magnetic capture probes by modifying oligonucleotides complementary to target sequences on the surface of amino-modified silica-coated magnetic nanoparticles and optimized the conditions and parameters of probe synthesis and hybridization. We innovatively put the complexes of magnetic capture probes and target sequences into qPCR without any need for denaturation and purification steps. This strategy can reduce manual steps and save time. We used the magnetic capture probes to separate invA mRNA from Salmonella in artificially contaminated milk samples. The detection sensitivity was 10(4) CFU/ml, which could be increased to 10 CFU/ml after a 12 h enrichment step. The developed method is robust enough to detect live bacteria in a complex environmental matrix.
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