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基于荧光标记和核酸适配子识别可卡因,建立了简单、灵敏的可卡因新型荧光分析法。在微孔板表面组装亲和素-生物素化可卡因适配子-FAM标记可卡因适配子互补短链复合物,根据加入可卡因前后荧光强度的变化来定量可卡因。实验考察了微孔板包被亲和素浓度、生物素标记适配子用量、FAM标记可卡因适配子互补短链用量、反应温度、反应时间等因素对体系组装和可卡因识别的影响。结果显示,实验选定条件下,可卡因浓度在5～1600 nmol/L范围内与荧光强度呈线性关系,检出限为3.0 nmol/L(3δ);尿样加标回收率为89.5%～98.6%,相对标准偏差小于5%。
A simple and sensitive new fluorescence analysis method for cocaine based on fluorescent labeling and aptamer recognition of cocaine was established. Assemble avidin-biotinylated cocaine aptamer-FAM-labeled cocaine aptamer complementary short-chain complex on the surface of microplate, and quantify the cocaine according to the change of fluorescence intensity before and after adding cocaine. The effect of factors such as the concentration of avidin in the microplate, the amount of biotin labeled aptamer, the complementary short-chain amount of FAM labeled cocaine aptamer, the reaction temperature and the reaction time on the assembly of the system and cocaine were investigated. The results showed that under the selected experimental conditions, the concentration of cocaine in the range of 5 ~ 1600 nmol / L showed a linear relationship with the fluorescence intensity, with a detection limit of 3.0 nmol / L (3δ). The spiked recoveries of urine samples ranged from 89.5% to 98.6 %, Relative standard deviation less than 5%.