Université Paris-Saclay, INRAE, AgroParisTech, Micalis Institute, 78350 Jouy-en-Josas, France.
Université Paris-Saclay, INRAE, GABI, 78350 Jouy-en-Josas, France.
J Microbiol Methods. 2022 Oct;201:106578. doi: 10.1016/j.mimet.2022.106578. Epub 2022 Sep 13.
Nonspecific binding of proteins from complex food matrices is a significant challenge associated with a biosensor using gold nanoparticles (AuNPs). To overcome this, we developed an efficient EDTA chelating treatment to denature milk proteins and prevent their adsorption on AuNPs. The use of EDTA to solubilize proteins enabled a sensitive label-free apta-sensor platform for colorimetric detection of Staphylococcus aureus in milk and infant formula. In the assay, S. aureus depleted aptamers from the test solution, and the reduction of aptamers enabled aggregation of AuNPs upon salt addition, a process characterized by a color change from red to purple. Under optimized conditions, S. aureus could be visually detected within 30 min with the detection limit of 7.5 × 10 CFU/mL and 8.4 × 10 CFU/mL in milk and infant formula, respectively. The EDTA treatment provides new opportunities for monitoring milk contamination and may prove valuable for biosensor point-of-need applications.
从复杂的食物基质中提取蛋白质时,金纳米粒子(AuNPs)生物传感器会遇到非特异性结合的问题。为了解决这个问题,我们开发了一种有效的 EDTA 螯合处理方法,使牛奶蛋白变性,防止其吸附在 AuNPs 上。使用 EDTA 溶解蛋白质,使基于比色检测的无标记适体传感器平台能够灵敏地检测牛奶和婴儿配方奶粉中的金黄色葡萄球菌。在该测定中,金黄色葡萄球菌从测试溶液中耗尽适体,并且由于盐的加入减少了适体,AuNPs 的聚集得以实现,这一过程的特征是颜色从红色变为紫色。在优化条件下,金黄色葡萄球菌可在 30 分钟内被视觉检测到,在牛奶和婴儿配方奶粉中的检测限分别为 7.5×10 CFU/mL 和 8.4×10 CFU/mL。EDTA 处理为监测牛奶污染提供了新的机会,并且可能对生物传感器即时应用具有重要价值。