摘要
该文基于适配体以及非巯基化核酸修饰的胶体金纳米探针(AuNPs@polyA-DNA),建立了一种新型的卡那霉素胶体金侧向层析试纸条。分析了试纸条各组装元件,包括适配体浓度、链霉亲和素(SA)与Biotin-DNAT的比例、SA-生物素(Biotin)-DNAT偶联物浓度以及孵育时间与温度等,对显色反应的影响。最佳实验条件为:缓冲液为4xSSC(0.5%Tween 20),SA与Biotin-DNAT的最佳摩尔比为1∶6,检测区喷涂偶联物SA-Biotin-DNAT浓度为4μmol/L,适配体浓度为10 nmol/L,室温孵育时间为20 min。在优化条件下,该试纸条对卡那霉素的肉眼分辨浓度为25 ng/mL,线性范围为5.0~125 ng/mL,检出限为1.5 ng/mL。用于蜂蜜中卡那霉素的检测,其回收率为95.1%~105%,相对标准偏差(RSD)为3.4%~8.5%。该试纸条具有灵敏度高、特异性好、架构简单、重复性高等优点,可用于实际样品中卡那霉素的检测。
In this paper,a new type of colloidal gold lateral flow test strip(AuNPs@polyA-DNA)was fabricated based on aptamer recognition of kanamycin molecule and non-thiolated DNA modified colloidal gold nanoprobe.Effects of the strip components on color development were investigated,including concentration of aptamer,proportion of streptavidin(SA)to Biotin-DNAT,concentration of SA-Biotin-DNAT,conjugate reaction time and temperature.The optimized experimental conditions were as follows:Running buffer:4xSSC(0.5%Tween 20),molar ratio of SA to Biotin-DNAT:1∶6,concentartion of SA-Biotin-DNAT conjugate sprayed at T line:4μmol/L,concentartion of aptamer:10 nmol/L,incubation of aptamer:ambient temperature for 20 min.The results demonstrated that 25 ng/mL kanamycin could be discriminated with the naked eyes.The quantitative determination of kanamycin in the range of 5.0-125 ng/mL could be carried out by a colloidal gold reader,with a detection limit of 1.5 ng/mL.The test strip was applied to the determination of kanamycin in honey,with recoveries of 95.1%-105%and relative standards deviation(RSD)of 3.4%-8.5%.The strip could be used for the detection of kanamycin in real samples with the advantages of high sensitivity,high specificity,simple structure and high repeatability.
作者
常芮
蔡伊娜
王周平
彭池方
CHANG Rui;CAI Yi-na;WANG Zhou-ping;PENG Chi-fang(State Key Laboratory of Food Science and Technology,Jiangnan University,Wuxi 214122,China;School of Food Science and Technology,Jiangnan University,Wuxi 214122,China;Food Inspection and Quarantine Centre,Shenzhen Customs,Shenzhen 518045,China)
出处
《分析测试学报》
CAS
CSCD
北大核心
2021年第12期1728-1735,共8页
Journal of Instrumental Analysis
基金
国家重点研发计划项目(2019YFC1605405)
国家自然科学基金项目(31371767)。