Gui Yun, Zhao Yun, Liu Pengyan, Wang Yulong, Mao Xinxin, Peng Chifang, Hammock Bruce D, Zhang Cunzheng
State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, P. R. China.
Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, P. R. China.
ACS Omega. 2024 Feb 2;9(6):7075-7084. doi: 10.1021/acsomega.3c09050. eCollection 2024 Feb 13.
Later flow immunochromatographic assay has been widely used in clinical, environmental, and other diagnostic applications owing to its high sensitivity and throughput. However, most immunoassays operate in the "turn-off" mode for detecting targets of low molecular weight. The signal intensity decreases as the analyte concentration increases, which poses a challenge for achieving ultrasensitive detection at low concentrations and is counterintuitive to new users. In this work, a fluorometric immunochromatographic assay (FICA) is developed to simultaneously read "turn-on" fluorescent and "turn-off" colorimetric signals, where ZnCdSe/ZnS quantum dots act as fluorescence donors and gold nanoparticles (AuNPs) act as quenchers. The fluorescent signal (excitation/emission wavelengths of 365/525 nm) is positively correlated with analytes' concentration. Taking sibutramine (SBT) as the analysis target, the visual limit of detection for SBT reached 3.9 ng/mL, and the limit of Quantitation was 5.0 ng/mg in spiked samples. The developed FICA achieves a high sensitivity in SBT detection, which is much lower than that of the colloidal gold-based immunochromatographic assay. This dual-function detection mode has great potential to be used as a rapid on-site semiquantitative method, providing an alternative mode for the determination of low levels of target analytes.
后来,流动免疫色谱分析法因其高灵敏度和高通量而被广泛应用于临床、环境及其他诊断应用中。然而,大多数免疫测定法在检测低分子量目标物时采用“关闭”模式。随着分析物浓度的增加,信号强度会降低,这给在低浓度下实现超灵敏检测带来了挑战,并且对新用户来说有违直觉。在这项工作中,开发了一种荧光免疫色谱分析法(FICA),以同时读取“开启”荧光信号和“关闭”比色信号,其中ZnCdSe/ZnS量子点作为荧光供体,金纳米颗粒(AuNPs)作为猝灭剂。荧光信号(激发/发射波长为365/525 nm)与分析物浓度呈正相关。以西布曲明(SBT)为分析目标,SBT的目视检测限达到3.9 ng/mL,加标样品中的定量限为5.0 ng/mg。所开发的FICA在SBT检测中实现了高灵敏度,远低于基于胶体金的免疫色谱分析法。这种双功能检测模式有很大潜力用作快速现场半定量方法,为低水平目标分析物的测定提供了一种替代模式。