快速检测生鲜乳中玉米赤霉醇试纸条的研制及其应用

Research and Application of Dipstick Detecting Zeranol in Raw Milk

[目的]采用免疫胶体金技术,建立了一种快速检测玉米赤霉醇的试纸条方法。[方法]利用有机合成方法,研制了一种玉米赤霉醇半抗原,将其载体蛋白偶联得到免疫抗原,并通过人工免疫的方法获得单克隆抗体。通过酶联免疫法对制备的单克隆抗体的性能进行了验证。玉米赤霉醇层析试纸条是利用单克隆抗体标记胶体金,其原理是玉米赤霉醇抗原和羊抗鼠Ig G分别喷到自制的膜(硝酸纤维)上,然后将样品垫、吸水垫分别放置在PVC板上,再制成试纸条。[结果]试纸条检测限可达0.5~1.0μg/kg,具有速度快、简单等优点。此外,该试纸条与大部分的玉米赤霉醇没有明显的交叉反应。[结论]利用该试纸条可实现对生鲜乳中玉米赤霉醇添加试验的准确判定。该试纸条有望实现对生鲜乳中玉米赤霉醇残留的快速筛查。

[Objective] The research aimed to establish a rapid strip detection method for zeranol by immune gold colloid technology. [Method]The hapten of zeranol was prepared via a synthetic organic method,and was conjugated with the carrier protein to obtain the antigen. Monoclonal antibody was obtained with the artificial immune method and its affinity was verified with the enzyme immunoassay. The colloid gold immunochromatographic strip was prepared with monoclonal antibody conjugated gold colloid which was stored in microwells,and a PVC liner which was assembled with nitrocellulose membrane sprayed with zeranol antigen and goat anti-mouse Ig G,sample pad and absorbent pad. [Result]The strip had a detection limit in the range of 0. 5-1. 0 μg/kg with the advantages of fastness,easiness,etc. In addition,the strip did not cross-react with most of the pyrethroid pesticides tested. [Conclusion]With the strip,the accurate determination of the addition of zearalan in raw milk is achieved. This strip is expected to achieve fast screening of zearalenol residues in raw milk.