基于纳米金标记的金黄色葡萄球菌可视化检测方法研究  

A nano-gold labling-based visual detection method for Staphylococcus aureus

作　　者：袁京磊 俞晔[2] 李灿[1] 王周平[1] 

机构地区：[1]江南大学食品学院,无锡214122 [2]张家港出入境检验检疫局,张家港215600

出　　处：《食品安全质量检测学报》2013年第4期1100-1108,共9页Journal of Food Safety and Quality

基　　金：国家"十二五"科技支撑计划项目(2012BAK08B01);江苏省科技支撑计划项目(BE2011621;BE2010679);教育部"新世纪优秀人才支持计划"(NCET-11-0663)~~

摘　　要：目的以纳米金为载体,对金黄色葡萄球菌进行可视化检测。方法将与金黄色葡萄球菌目标序列互补的DNA1和DNA2连接到纳米金上,当体系中出现金黄色葡萄球菌目标序列时,两条短链DNA就会与目标序列杂交,使纳米金之间的距离拉近,从而使纳米金发生团聚,导致纳米金的颜色从酒红色变为蓝色。由于金黄色葡萄球菌目标序列浓度的不同,从而引起纳米金之间团聚的程度不同,纳米金就会相应的呈现出不同的颜色变化,这样就可达到可视化检测的目的。结果在最优实验条件下,本方法在检测金黄色葡萄球菌目标序列时,浓度在1～1000pmol/L范围内呈现良好的线性关系,检出限为0.5pmol/L;检测金黄色葡萄球菌时,线性范围为30～9800CFU/mL,检出限为25CFU/mL。结论通过特异性和加标回收实验,证明本方法可以用于实际样品的检测。Objective To set up a visual detection method of Staphylococcus aureus based on the col-or-changing effect of AuNPs. Methods The DNA 1 and DNA 2 which were complementary with the target DNA of Staphylococcus aureus were connected to the surface of AuNPs. When the target sequence of Staphy-lococcus aureus appeared, the two short-chain DNAs hybridized to the target sequence, and the AuNPs became closer, then the color of AuNPs turned blue from wine red. The different concentrations of target sequence caused different degrees of aggregation between AuNPs, and the AuNPs accordingly showed different colors . Results Under optimal conditions, the correlation between concentration of target sequence of Staphylococ-cus aureus and A700/A525 was observed to be linear within the range of 1~1000 pmol/L, and the detection limit of the proposed method was 0.5 pmol/L. The correlation between concentration of Staphylococcus aureus and A700/A525 was observed to be linear within the range of 30~9800 CFU/mL, and the detection limit of the proposed method was 25 CFU/mL. Conclusion The specificity and spiked recovery experiments proved that this developed method could be used for detection of actual sample.

关 键 词：纳米金 金黄色葡萄球菌 可视化检测 

分 类 号：R378[医药卫生—病原生物学]