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基于光激化学发光均相免疫检测技术的黄曲霉毒素检测方法研究

Study of the aflatoxin detection method based on luminescent oxygen channeling immunoassay technology

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摘要

黄曲霉毒素是一种毒性极强的生物毒素,广泛存在于各种农产品与食品中,对公众健康危害极大。提出了一种基于光激化学发光均相免疫检测技术的黄曲毒素B1检测方法,利用生物素亲和及抗原与抗体免疫反应,形成受体微球-AFB1-BSA完全抗原-AFB1单抗体-供体微球的聚合体系,在激发光作用下产生化学发光反应,根据化学发光值可定量分析样品中黄曲霉毒素浓度。通过分析并优化反应条件,包括各种反应物浓度、反应体系体积、反应温度与时间等,可实现最低检测限0.048ng/mL、最低定量限0.22ng/mL、批内与批间变异系数均小于10%、植物油样品中AFB1的加标回收率在87~105%之间。方法灵敏度高、特异性强、免清洗、检测速度快,在真菌毒素现场快速检测领域具有广阔的应用前景。

Abstract

Aflatoxin is a highly toxic biological toxin that is widely existed in agricultural products and food. It endangers public health. In the manuscript, a method based on luminescent oxygen channeling immunoassay technology is used for aflatoxin B1 detection. By using the affinity of biotin and the immune reaction between the antigen and the antibody, a polymer system of acceptor beads-AFB1-BSA complete antigen-AFB1 monoclonal antibody-donor beads is formed. Then, with the excitation of the laser, the chemiluminescence reaction produces excitation light. The aflatoxin concentrations could be calculated by the intensity of excitation light. The experimental conditions are analyzed and optimized, including the volume, the concentration of various reagents, incubation temperature and time. However, the method can achieve the lower detection limit of 0.048ng/mL, the lower limit of quantification of 0.22ng/mL, the intra- and inter-assay variation both below 10%, and the recovery rates 87~105% with oil samples. Taking the advantages of high sensitivity, strong specificity, and wash-free, this method could be a promising tool for rapid detection of toxins.

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所属栏目:生物光学

基金项目:国家自然科学基金(81401454,81471698,81671727); 中国博士后科学基金(2013M541174); 中国博士后国际交流计划(20140066); 天津市应用基础与前沿技术研究计划 (16JCZDJC31200)

收稿日期:2017-09-28

修改稿日期:2017-12-22

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吴震:天津大学 精密仪器与光电子工程学院, 天津 300072
宗婧:天津大学 精密仪器与光电子工程学院, 天津 300072
李晨曦:天津大学 精密仪器与光电子工程学院, 天津 300072
陈文亮:天津大学 精密仪器与光电子工程学院, 天津 300072天津大学 精密测试技术及仪器国家重点实验室, 天津 300372
徐可欣:天津大学 精密仪器与光电子工程学院, 天津 300072天津大学 精密测试技术及仪器国家重点实验室, 天津 300372

联系人作者:吴震(zhenwu@tju.edu.cn)

备注:吴震(1990-),男,硕士研究生,从事均相免疫检测技术、食品安全检测研究。

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引用该论文

WU Zhen,ZONG Jing,LI Chenxi,CHEN Wenliang,XU Kexin. Study of the aflatoxin detection method based on luminescent oxygen channeling immunoassay technology[J]. Optical Technique, 2019, 45(1): 117-123

吴震,宗婧,李晨曦,陈文亮,徐可欣. 基于光激化学发光均相免疫检测技术的黄曲霉毒素检测方法研究[J]. 光学技术, 2019, 45(1): 117-123

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