State Key Laboratory of Food Science and Technology, Synergetic Innovation Center of Food Safety and Nutrition, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.
School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, China.
Talanta. 2017 Mar 1;164:275-282. doi: 10.1016/j.talanta.2016.11.053. Epub 2016 Nov 23.
A sensitive steady-state chemiluminescent aptasensor based on rolling circle amplification (RCA) was fabricated for the detection of Salmonella typhimurium. The sensor utilized aptamer modified FeO magnetic nanoparticles (MNPs) as capture probes, aptamer as recognition molecules, and Co enhanced N‑(aminobutyl)-N-(ethylisoluminol) (ABEI) functional flowerlike gold nanoparticles (AuNFs) and complementary strand (cDNA) complex (Co/ABEI-AuNFs-cDNA) as signal probes. And P-Iodophenol (PIP) was also added to form a typical ABEI- AuNFs-PIP-HO steady-state CL system. By virtue of FeO MNPs based solid-phase RCA strategy, S. typhimurium can be first captured by the aptamer immobilized on the surface of FeO MNPs then complex with RCA products to form a sandwich complex. Co/ABEI-AuNFs-cDNA signal probes were then assembled on the RCA products to produce and enhance CL signals. Under optimal conditions, the logarithmic correlation between the concentration of S. typhimurium and the CL signal was found to be linear within the range of 32cfumL to 3.2×10cfumL (R =0.9921). The limits of detection of the developed method were found to be 10cfumL for S. typhimurium. The method was also used to detect S. typhimurium in real pork samples. The results were compared with those obtained from the plate-counting methods and showed good consistency. Therefore, this detection aptasnesor can be a good candidate for sensitive and selective detection of S. typhimurium with simple and effective operations.
基于滚环扩增（RCA）的灵敏稳态化学发光适体传感器被制备用于检测鼠伤寒沙门氏菌。该传感器利用适配体修饰的 FeO 磁性纳米粒子（MNPs）作为捕获探针，适配体作为识别分子，以及 Co 增强的 N-（氨丁基）-N-（乙基异鲁米诺）（ABEI）功能花状金纳米粒子（AuNFs）和互补链（cDNA）复合物（Co/ABEI-AuNFs-cDNA）作为信号探针。此外，还添加了 P-碘苯酚（PIP）以形成典型的 ABEI-AuNFs-PIP-HO 稳态 CL 系统。基于 FeO MNPs 的固相 RCA 策略，鼠伤寒沙门氏菌首先可以被固定在 FeO MNPs 表面的适配体捕获，然后与 RCA 产物复合形成三明治复合物。然后，Co/ABEI-AuNFs-cDNA 信号探针被组装在 RCA 产物上以产生和增强 CL 信号。在最佳条件下，发现鼠伤寒沙门氏菌浓度与 CL 信号之间的对数相关性在 32cfumL 至 3.2×10cfumL（R = 0.9921）范围内呈线性关系。该方法的检测限被发现为 10cfumL 的鼠伤寒沙门氏菌。该方法还用于检测实际猪肉样品中的鼠伤寒沙门氏菌。结果与平板计数法的结果进行了比较，显示出良好的一致性。因此，这种检测适体传感器可以作为一种灵敏、选择性检测鼠伤寒沙门氏菌的良好候选物，具有简单有效的操作。
