Lian Fengnan, Wang Dan, Yao Shuo, Ge Lirui, Wang Yue, Zhao Yuyi, Zhao Jinbin, Song Xiuling, Zhao Chao, Li Jinhua, Liu Yajuan, Jin Minghua, Xu Kun
School of Public Health, Jilin University, 130021 Changchun, China.
Jilin Engineering Research Center of Public Health Detection, 130021 Changchun, China.
Food Sci Biotechnol. 2021 Jul 28;30(8):1129-1138. doi: 10.1007/s10068-021-00947-3. eCollection 2021 Aug.
This research aimed to detect O157:H7 in milk based on immunomagnetic probe separation technology and quenching effect of gold nanoparticles to Rhodamine B. Streptavidin-modified magnetic beads (MBs) were combined with biotin-modified antibodies to capture O157:H7 specifically. Gold nanoparticle (AuNPs) was incubated with sulfhydryl-modified aptamers (SH-Aptamers) to obtain the Aptamers-AuNPs probe. After magnetic beads captured target bacteria and formed a sandwich structure with the gold nanoprobe, Rhodamine B was added into complex to obtain fluorescent signal changes. Our results demonstrated that the established method could detect O157:H7 in the range of 10-10 CFU/mL, and the limit of detection (LOD) was 0.35 CFU/mL in TBST buffer (pH = 7.4). In milk simulation samples, the LOD of this method was 1.03 CFU/mL. Our research provides a promising approach on the detection of O157:H7.
本研究旨在基于免疫磁珠分离技术及金纳米粒子对罗丹明B的猝灭作用检测牛奶中的O157:H7。链霉亲和素修饰的磁珠(MBs)与生物素修饰的抗体相结合,特异性捕获O157:H7。金纳米粒子(AuNPs)与巯基修饰的适配体(SH-适配体)孵育,得到适配体-金纳米粒子探针。磁珠捕获目标细菌并与金纳米探针形成夹心结构后,向复合物中加入罗丹明B以获得荧光信号变化。我们的结果表明,所建立的方法能够在10-10 CFU/mL范围内检测O157:H7,在TBST缓冲液(pH = 7.4)中的检测限(LOD)为0.35 CFU/mL。在牛奶模拟样品中,该方法的LOD为1.03 CFU/mL。我们的研究为O157:H7的检测提供了一种有前景的方法。