免疫快速同步检测玉米中5种真菌毒素胶体金试纸条的构建和应用

Development and Application of Colloidal Gold Test Strip for Simultaneously and Fast Detection of Five Mycotoxins in Maize

玉米作为重要的粮食和饲料来源,容易受到多种真菌毒素的污染,给居民健康和养殖业造成严重的危害。本文优化了适合5种真菌毒素(黄曲霉毒素B1、伏马毒素、T-2毒素、脱氧雪腐镰刀菌烯醇和玉米赤霉烯酮)同时提取的样品前处理方法,并构建了5检测线胶体金试纸条,检测结果可以通过肉眼定性,结合Image J软件可以进行定量分析,并应用于实际玉米样品检测。结果表明,最佳提取条件为90%乙腈/水,涡旋20 min,样品的添加回收率为71.9%~113.3%,相对标准偏差为0.9%~7.5%;测定基质添加样品建立标准曲线,通过肉眼判定得到5种真菌毒素的消线值分别为80.0、1 000.0、100.0、400.0、200.0μg/kg;结合Image J软件提取多重检测试纸条的检测线灰度值,结果表明当AFB1、FB1、T-2、DON和ZEN的质量分数为8.0、20.0、10.0、40.0、20.0μg/kg时,各真菌毒素浓度对应的检测线显色值比空白对照显色值低。最后,通过检测11个实际玉米样品,结果表明部分玉米样品受到了不同程度的AFB1、DON和ZEN的污染。因此,本研究开发的免疫快速同步检测玉米中5种真菌毒素胶体金试纸条可以用于玉米中真菌毒素的现场快速筛查。

As an important crop and feed source,maize is easily contaminated by a variety of mycotoxins,causing serious harm to residents'health and aquaculture.In this paper,a pretreatment method suitable for the simultaneous extraction of five mycotoxins(aflatoxin B 1,fumonisin,T-2 toxin,deoxynivalenol and zearalenone)was optimized,and colloidal gold test strip with multiple detection lines was constructed,which could be qualitatively analyzed by the naked eye.Combined with Image J software,the results could also be analyzed quantitatively.This method was applied to detect the actual corn sample.The results indicated that the samples could be extracted by 90%acetonitrile/water for 20 min with 71.9%~113.3%recovery rate and 0.9%~7.5%the relative standard deviation.The standard curves were determined,and the cutoff values of the five mycotoxins were 80.0,1000.0,100.0,400.0 and 200.0μg/kg,respectively.Combined with Image J software,the extracted gray area of the detection line of the multiplex test strip showed that when the concentrations of AFB 1,FB 1,T-2,DON and ZEN were 8.0,20.0,10.0,40.0,20.0μg/kg,the color intensity was lighter than that of the blank control.Finally,the results indicated that some maize samples are contaminated by AFB 1,DON and ZEN with different pollution level.Therefore,the developed colloidal gold test strip could be used for on-site rapid screening of five mycotoxins in maize.