Homogeneous time-resolved FRET assay for the detection of Salmonella typhimurium using aptamer-modified NaYF4: Ce/Tb nanoparticles and a fluorescent DNA label
Homogeneous time-resolved FRET assay for the detection of Salmonella typhimurium using aptamer-modified NaYF4: Ce/Tb nanoparticles and a fluorescent DNA label
复制标题
使用适体修饰的 NaYF4:Ce/Tb 纳米颗粒和荧光 DNA 标记进行均相时间分辨 FRET 测定，用于检测鼠伤寒沙门氏菌
DOI：
10.1007/s00604-017-2399-5
复制
发表时间：
2017
影响因子：
5.7
通讯作者：
Wang Zhouping
中科院分区：
文献类型：
--
作者：
Wang Xiaole;Niazi Sobia;Yukun Huang;Sun Weijia;Wu Shijia;Duan Nuo;Hun Xu;Wang Zhouping
作者描述了一种基于时间分辨荧光共振能量转移（TR-FRET）的适体传感器，用于鉴定鼠伤寒沙门氏菌（S。鼠伤寒沙门氏菌）。适配体功能化的纳米粒子（NP）被用作能量供体，和互补的寡核苷酸（cDNA）标记的羧基荧光素（FAM）作为受体。该检测方案基于适体和cDNA之间的杂交，在此基础上，光子能量从NP转移到FAM，除非适体与S相互作用。鼠伤寒。由于核酸适体的高度特异性识别能力和纳米粒的强荧光强度，该方法对S具有高度的灵敏度和选择性。鼠伤寒。在最佳条件下，激发波长为273 nm，延迟时间为100 μs，门控时间为1 ms，积分时间分辨荧光强度比（FAM 520/Tb 489）在100 ~ 106 cfu·mL-1范围内呈线性关系，检测限低至25 cfu·mL-1。该方法已应用于鸡蛋和鸡肉中S。图形摘要通过NaYF 4：Ce/Tb纳米粒子标记的适体和FAM-cDNA之间的FRET，用于测定鼠伤寒沙门氏菌的均相时间分辨FRET分析的示意图，具有良好的线性和25 cfu mL−1的检测限。
The authors describe an aptasensor based on time-resolved fluorescence resonance energy transfer (TR-FRET) for the identification ofSalmonella typhimurium(S. typhimurium). Aptamer-functionalized nanoparticles (NPs) is used as the energy donor, and a complementary oligonucleotide (cDNA) labeled with carboxyfluorescein (FAM) acts as the acceptor. The detection scheme is based on the hybridization between aptamer and cDNA, upon which photonic energy is transferred from NPs to FAM unless aptamer interacts withS. typhimurium. Due to the highly specific recognition ability of aptamer and the strong fluorescence intensity of NPs, the method is highly sensitive and selective forS. typhimurium. Under the optimal conditions, at excitation wavelength of 273 nm, a delay time of 100 μs and a gating time of 1 ms, the integrated time-resolved fluorescence intensity ratio (FAM520/Tb489) is linear in the 100 to 106cfu·mL−1range, and the limit of detection is as low as 25 cfu·mL−1. The assay was applied to the analysis of eggs and chicken meat forS. typhimurium, and the results were consistent with those of a plate-counting method.Graphical abstractSchematic of a homogeneous time-resolved FRET assay for the determination ofSalmonella typhimuriumby FRET between NaYF4:Ce/Tb nanoparticle-tagged aptamer and FAM-cDNA, with a good linearity and the limit of detection of 25 cfu mL−1.