量子点荧光微球免疫层析试纸条定量检测玉米中赭曲霉毒素A  

Immunochromatographic Assay for Quantitative Detection of Ochratoxin A in Maize by Quantum Dot Beads

作　　者：周耀锋 熊斯诚 江湖[1,2] 段宏[1,2] 熊勇华[1,2] Andrew Wang 

机构地区：[1]食品科学与技术国家重点实验室,南昌大学,南昌330047 [2]中德联合研究院,南昌大学,南昌330047 [3]Ocean NanoTech, LLC. , San Diego, CA92126 USA

出　　处：《分析化学》2015年第12期1837-1843,共7页Chinese Journal of Analytical Chemistry

基　　金：国家重点基础研究发展计划(973计划,No.2013CB127804);国家自然科学基金(No.31271863);江西省主要学科与技术带头人培养计划(No.2014BCB22004)资助~~

摘　　要：采用1-乙基-(3-二甲基氨基丙基)碳二亚胺(EDC)法将抗赭曲霉毒素A(OTA)腹水与量子点荧光微球偶联制备荧光探针,以牛血清白蛋白-OTA偶联物及驴抗鼠二抗喷涂硝酸纤维膜形成试纸条检测线(T线)和质控线(C线),建立了基于T/C荧光强度(FI_T/FI_C)比值法免疫层析试纸条高灵敏度、快速定量检测玉米中OTA的新方法。量子点荧光微球试纸条定量检测OTA线性范围为0.05~1.0ng/mL(Y=0.259lnX+0.897,R^2=0.995),半数抑制浓度(IC_(50))为(0.215±0.023)ng/mL(n=5),玉米提取液中OTA的检出限为0.05 ng/mL,单个样品检测时间10min。加标回收实验表明,3个OTA加标浓度的平均回收率为107.2%-125.5%,相对标准偏差均小于10%。40个实际玉米样品检测结果表明,量子点荧光微球试纸条与酶联免疫吸附分析方法检测OTA的结果具有良好的相关性(R^2=0.975)。Fluorescence probes were prepared by coupling the ascetic fluid containing anti-ochratoxin A（OTA） monoclonal antibody with quantum dot beads（QBs） using 1-（3-dimethylaminopropyl）-3—ethylcarbodiimide hydrochloride method.The OTA-labeled bovine serum albumin（BSA） conjugates and donkey anti-mouse polyclonal antibodies were sprayed onto the nitrocellulose membrane as the test and control lines,respectively.The resultant fluorescence probes were introduced into the immunochromatographic strip（ICA） for sensitive,rapid and quantitative detection of OTA in maize based on the ratio of fluorescence intensities of test and control lines.The QBs-based ICA exhibited dynamic linear range for detection of OTA in maize extract from 0.05 ng/mL to 1.0 ng/mL（Y = 0.2591 nX ＋ 0.897,R^2 = 0.995）,with the median inhibitory concentration of（0.215 ±0.023） ng/mL（n =5）.The detection limit for OTA was 0.05 ng/mL in maize extract,and the detection time of the proposed QBs-based ICA for each sample was 10 min.The recoveries of OTA at three spiked levels were 107.2%- 125.5%,while relative standard deviations were below 10%.Forty natural maize samples were assayed using both QB-ICA and enzyme-linked immunosorbent assay,and the results from the two methods showed a highly significant correlation（R = 0.975）.

关 键 词：量子点荧光微球 免疫层析试纸条 赭曲霉毒素A 定量检测 

分 类 号：TS207[轻工技术与工程—食品科学] O657.3[轻工技术与工程—食品科学与工程]