摘要
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,sensitive,and rapid fluorescent analytical method was developed based upon fluorescent labeling and aptamer recognition.The complex of avidin-biotinylated cocaine aptamer-FAM labeled part-complementary strand of cocaine aptamer was assembled onto the microplate wells.Cocaine was quantified according to the change of the fluorescent intensity induced by the dissociated FAM labeled part-complementary strand of cocaine aptamer.Factors including the amount of avidin coated onto microplate wells,biotinylated cocaine aptamer,FAM labeled part-complementary strand of cocaine aptamer,reaction temperature and time,that affect the assembly and recognition process were investigated.The results show that under the selected conditions,the concentration of cocaine is linear to the fluorescent intensity ranged from 5 to 1600 nmol/L with a detection limit 3.0 nmol/L（3S/N）.The standard recoveries were 89.5%~98.6% for urine samples and the RSD is below 5%.
出处
《分析试验室》
CAS
CSCD
北大核心
2011年第4期19-21,共3页
Chinese Journal of Analysis Laboratory
基金
"863"计划(2008AA10Z419)
江苏省自然科学基金(BK20081603)项目资助
