摘要
[目的]建立用于快速检测伏马菌素B1(FB1)的胶体金免疫层析试纸条。[方法]首先制备了FB1单克隆抗体和FB1-OVA偶联抗原,然后将FB1单克隆抗体与胶体金制成金标抗体包被在金标垫上,将FB1-OVA抗原和羊抗鼠Ig G包被在硝酸纤维素膜(NC膜)表面分别作为检测线和质控线。采用间接竞争法检测待检样品中的FB1与NC膜上的FB1-OVA抗原竞争结合金标抗体中的FB1单克隆抗体情况,并以检测线和质控线显示检测结果。[结果]本试验优化了金标抗体、检测抗原及羊抗鼠Ig G的包被量,检测FB1的灵敏度可达到2 ng·m L-1,最佳判定质量浓度为40 ng·m L-1。[结论]本试验制备的检测试纸条具有较高的特异性和稳定性,操作简单,检测时间仅需10 min,且与高效液相色谱法检测结果一致,可作为检测玉米中伏马菌素B1残留的有效手段。
[ Objectives] A rapid golden immunochromatographic strip for the detection of fumonisin B1（FB1）was developed. [Methods] Specific monoclonal antibody against FB1 and FB：OVA conjugate antigen was prepared. FB1 monoclonal antibody, labeled with colloidal gold,was used as probes on the immunochromatographic strip. FB,-OVA and goat anti-mouse IgG were coated onto nitrocellulose membrane as test lines and control lines,respectively. FB, in sample will competitively combined the FB, mono- clonal antibody with the FB1-OVA in nitrocellulose filter membrance（NC） and the result will be observed by the color of detection line and quality control line directly. [ Results ] In this study, the concentrations of FB1 monoclonal antibody labeled with colloidal gold,detecting antigen and goat anti-mouse IgG were optimized. During the detection of corn, the lowest detectable limit reaches 2 ng·mL^-1 ,and the best determination concentration reaches 40 ng·mL^-1. The test can be completed only within 10 min. [ Conclusions] The detection of samples showed that the developed method had a positive correlation with the HPLC method. It can be an effective method to detect FB1 in corns.
出处
《南京农业大学学报》
CAS
CSCD
北大核心
2015年第3期483-490,共8页
Journal of Nanjing Agricultural University
基金
安徽省科技攻关项目(12010402c206)
