摘要
为研制一种快速定量检测玉米中T-2毒素含量的胶体金试纸条,对T-2毒素单克隆抗体进行标记并对胶体金试纸条性能进行研究。用胶体金标记T-2毒素单克隆抗体,通过测定加入不同量碳酸钾后吸光度值的变化,确定标记的最佳pH值,通过与抗原进行正交试验确定最佳组合条件,应用胶体金定量读数仪通过检测线和对照线的对比,检测试纸条的性能。结果表明,单克隆抗体标记的最佳pH值为8.5,最佳标记浓度为20μg/mL,抗原的包被浓度为0.5mg/mL,羊抗鼠IgG的包被浓度为1.0mg/mL,试纸条检测T-2毒素标准品的检测限为5μg/kg。T-2毒素胶体金快速定量检测试纸条检测方法可得出具体数据,与常见真菌毒素无交叉反应,回收率在77%~117.6%之间,批内和批间重复性检测变异系数小于10%,玉米中T-2毒素的检测结果与高效液相法检测结果一致。T-2毒素胶体金快速定量检测试纸条可用于玉米中T-2毒素含量的快速定量检测。
In order to develop a rapid quantitative test strip for the detection of T-2 toxin in corn, the T-2 toxin monoclonal antibody was labeled and the performance of colloidal gold test strip was studied. The T-2 toxin monoclonal antibody was labeled with colloidal gold, and the optimum pH value of the label was determined by measuring the change of the absorbance value after adding different amounts of potassium carbonate. The optimal combination condition was determined by orthogonal experiment with the antigen. The performance of the strip was tested by comparing the detection line and the control line with the colloidal gold quantitative reader. The results showed that the optimal pH value of monoclonal antibody labeling was 8.5, the optimal labeling concentration was 20 μg/mL, the coating concentration of antigen was 0.5 mg/mL, and the coating concentration of goat anti-mouse IgG was 1.0 mg/mL. The detection limit of the strip test T-2 toxin standard was 5 μg/kg. T-2 toxin colloidal gold rapid quantitative test strip test method could obtain concrete data, and there was no cross-reaction with common mycotoxin, the recovery rate was between 77 % and 117.6 %, repetitive detection between batch and batch of variation coefficient was less than 10 %, and the detection result of T-2 toxin in corn was consistent with the results of high-performance liquid method. The T-2 toxin colloidal gold rapid quantitative test strip was successfully developed, which can be used for rapid quantitative detection of T-2 toxin content in corn.
作者
许会静
李玉杰
张馨月
王弘瑞
苗茁
柳玉
李艳
王会岩
董媛
XU Hui-jing;LI Yu-jie;ZHANG Xin-yue;WANG Hong-rui;MIAO Zhuo;LIU Yu;LI Yan;WANG Hui-yan;DONG Yuan(College of Laboratory Medicine, Jilin Medical College, Jilin 132013, Heilongjiang, China;Antibody Center, Jilin Medical College, Jilin 132013, Heilongjiang, China)
出处
《食品研究与开发》
CAS
北大核心
2019年第19期175-180,共6页
Food Research and Development
基金
国家科技重大专项(2012ZX09103-301-003)
吉林省教育厅“十二五”科学技术研究项目(吉教科合字[2015]第397号)
吉林省大学生创新训练项目(201731、201831)
关键词
T-2毒素
单克隆抗体
免疫层析
胶体金
快速检测
试纸条
T-2 toxin
monoclonal antibody
immunochromatography assay
colloidal gold
rapid detection
strip
