圆弧青霉菌毒素——青霉酸ELISA定量检测试剂盒的研制

The development of ELISA-kit of quantitative analysis for penicillic acid from Penicillium cyclopium

为了建立用于定量检测饲料与食品中青霉酸的ELISA试剂盒,试验在合成青霉酸人工抗原并免疫小鼠获得特异单克隆抗体的基础上,利用间接竞争ELISA方法对试剂盒各项技术参数进行测定,最终获得特异、快速的检测试剂盒。结果表明:在优化的条件下,该试剂盒对青霉酸标准品的最低检测浓度为1.6μg/mL;线性方程为y=9.54x+5.844 4(R2=0.996 4),线性范围为0.1～25.6μg/mL;回收试验的平均回收率分别为80.5%、83.3%、85.36%,37℃可保存48 h左右,与黄曲霉毒素B1、玉米赤霉烯酮、T-2毒素、伏马毒素等的交叉反应率小于0.01%,具有良好的特异性。说明已初步研制出用于检测饲料中青霉酸的ELISA试剂盒。

To develop initially the ELISA - kit used to detective quantitatively the penicillic acid in food and feed. On the basic of artificial antigen and monoclonal antibody required through immunizing the nfice,the technical parameters of the kit were determined bv indirect competitive ELISA,ultimately obtaining a special and rapid test kit. Under the optimized conditions,the limited concentration of kit was 1.6 μg/mL,linear equation was y = 9.54x ＋ 5. 844 4 （R^2 =0. 996 4） with linear range of 0.1 - 25.6 μg/mL. The recoveLy rate of kit was 80.5% ,83.3%, 85.36% respectively. The kit could be stored at 37℃about 48 hours. The cross - reaction rates with aflatoxin B1 ,zearalenone,T -2 toxin and fumonisins were less than 0.01% , and it showed that the kit had good specificity. The ELISA - kit used to detect penicillie acid in feed was developed initially.