氨基化磁性纳米粒子捕获基因组DNA结合PCR快速检测牛奶中金黄色葡萄球菌

Development of a rapid detection method for Staphylococcus aureus in milk based on magnetic separation using amino modified silica coated magnetic nanoparticles and polymerase chain reaction

摘要: 本研究对氨基化磁性纳米粒子进行优化,再用优化后的纳米粒子分离牛奶样品中微量的金黄色葡萄球菌基因组DNA,结合PCR检测,可以达到100 CFU/m L的检测限。在此过程中,纳米粒子与DNA的复合物直接加入到PCR体系中进行检测,无需洗脱、纯化等步骤,这样既减少了操作步骤,也减少了DNA损耗,还起到了浓缩的作用。而且,此方法中纳米粒子无需抗体等生物分子标记,成本非常低廉,在生命物质的快速检测中具有广阔的前景与巨大的潜力。

Abstract: In this study, the synthesis of amino-modified silica-coated magnetic nanoparticles were optimized, the optimal nanoparticles were used to adsorb a trace amount of genetic DNA from Staphylococcus aureus in milk, and then the complexes were used as DNA templates to be added into PCR.The limit of detection could reach 100 CFU/m L.The DNA and nanoparticle were directly added to PCR system during this process, without any steps of elution and purification. The cost of this magnetic separation method was quite low, for there was no need for nanoparticles to be labelled with biologic molecules such as antibody.This method has a great potential in the rapid detection of living organism.