金标记羟胺放大化学发光检测赭曲霉毒素A  

Detection of Ochratoxin A by Hydroxylamine-amplified Gold Nanoparticles Chemiluminescence

作　　者：李素 肖义陂[2] 武乐[3] 姜萌萌 刘杰[4] 严喜鸾 LI Su;XIAO Yi-bei;WU Le;JIANG Meng-meng;LIU Jie;YAN Xi-luan(School of Resources,Environmental&Chemical Engineering,Nanchang University,Nanchang 330031,China;Hongdu Traditional Chinese Medical Hospital,Nanchang 330008,China;3.Jiangxi Tumor Hospital, Nanchang 330029,China;Jiangxi Tumor Hospital, Nanchang 330029,China;School of Pharmacy,Nanchang University,Nanchang 330031,China)

机构地区：[1]南昌大学资源环境与化工学院,江西南昌330031 [2]南昌市洪都中医院,江西南昌330008 [3]江西省肿瘤医院,江西南昌330029 [4]南昌大学药学院,江西南昌330031

出　　处：《分析测试学报》2018年第1期57-61,共5页Journal of Instrumental Analysis

基　　金：国家自然科学基金项目(31660491);江西省自然科学基金项目(20132BAB205106)

摘　　要：以羧基磁性微球为分离载体,连接氨基捕获探针和适配体,加入生物素化报告序列和赭曲霉毒素A(Ochratoxin A,OTA)竞争结合适体,继续加入链霉亲和素纳米金和羟胺/Au^(3+)以显著提高化学发光检测OTA的灵敏度,从而建立了一种纳米金标记羟胺放大化学发光检测OTA的高灵敏度方法。优化了羧基磁性微球、氨基捕获探针、适配体、生物素化报告序列、链霉亲和素纳米金的用量。优化条件下,在OTA质量浓度0.01~50 ng/m L范围内,化学发光信号值与OTA浓度的对数呈较好的线性关系(r^2=0.992 5),检出限为1.58×10^(-3)ng/mL。对啤酒样品进行OTA加标回收实验,回收率为97.4%~105.4%,相对标准偏差为4.0%~5.5%。A novel chemiluminescent(CL)aptasensor for ochratoxin A(OTA)with magnetic beads as separate carrier was designed.Firstly,the capture DNA and OTA aptamer were immobilized on the surface of the magnetic beads,then the biotin-reporter DNA and OTA were taken to competitively bind to the OTA aptamer.Finally,streptavidin-gold nanoparticles(SA-AuNPs)and NH 2OH/Au 3+were added sequencely in order to improve the sensitivity for CL detection on OTA.The effects on the experiment including magnetic beads,capture DNA,aptamers,biotin-reporter DNA and dilution multiple of SA-AuNPs,were optimized.Under the optimized experimental conditions,there existed a good linear relationship between the CL intensity(ΔCL)and the logarithms of OTA concentration(r 2=0.992 5)in the range of 0.01-50 ng/mL,with a detection limit of 1.58×10-3 ng/mL.In the recovery experiments by adding different concentrations of OTA to the beer samples,the recoveries ranged from 97.4%to 105.4%with the relative standard deviations of 4.0%-5.5%.

关 键 词：赭曲霉毒素A 链霉亲和素纳米金 羟胺 化学发光 

分 类 号：O657.32[理学—分析化学] O433.4[理学—化学]