Elispot assay检测牛奶中庆大霉素

Establishment of Elispot Assay for Detection of Gentamicin in Milk

摘要:

为了建立快速检测牛奶中庆大霉素的Elispot assay,采用制备GM免疫抗原获得抗GM多克隆抗体。将检测抗原点阵在PVDF膜上,通过检测抗原和样品中GM竞争结合抗GM多克隆抗体,酶标结合物催化底物显色,根据颜色的有无及深浅判读结果,从而建立了检测牛奶中GM的Elispot assay。该方法的检测阈值为10 ng/mL,检测时间为40 min,可对样品实现半定量检测。该方法制备的试纸条于4℃密封保存90 d仍可用于检测;与多种结构类似物未见交叉反应;其结果与酶联免疫方法一致。

Abstract:

This manuscript developt a rapid detection method Elispot assay for Gentamicin sulfate,for this,Anti-GM polyclonal antibody through GM immunogen was obtained;Establishing milk GM-Elispot assay method by spotting coating antigen on PVDF membrane,and the coating antigen and GM in samples competitive binding the site of anti-GM polyclonal,then the HRP catalyzed chromogenic substrate,according to presence or absence of color and depth for interpretation of results.Results: The detection thre-shold of this method was 10 ng/mL,detection time was within 40 minutes and it can achieve semiquantitative detection.The strip of which was stored at 4 ℃ for 90 days could be used and no cross-reaction was found with several structural analogues.The results of samples analysis obtained from the Elispot assay were in a good agreement with that of ELISA method.