玉米赤霉烯酮ELISA定量检测试剂盒研制

目的研制玉米赤霉烯酮ELISA快速定量检测试剂盒,用以检测食品中抗玉米赤霉烯酮(ZEN)。方法在ZEN单克隆抗体基础上,用间接竞争酶联免疫吸附法研究试剂盒的各项指标。结果试剂盒最低检出浓度为1ng/ml,标准曲线的线性范围为1~200ng/ml,回归方程为^Y=0·99-0·40X(R2=0·99);抗体对ZEN50%的抑制浓度为16·3ng/ml;对玉米和小麦的平均加标回收率分别是96·5%和95·5%,变异系数分别是13·2%和10·9%;试剂盒4℃保质期超过6个月;对参试真菌毒素的交叉反应率均小于1%;实验室内的变异系数小于15%,实验室间变异系数小于20%;用本法与HPLC法同时检测美国维康公司玉米赤霉烯酮盲样,结果均在允许范围之内,且两种检测方法无差别。结论该试剂盒各项指标符合相关技术要求,具有快速、灵敏、稳定、专一的特点。

Development of ELISA-kit of quantitative analysis for Zearalenone

OBJECTIVE: To develope a rapid, sensitive, quantitative ELISA-kit for Zearalenone and determine zearalenone in cereals. METHODS: On the base of monoclonal antibodies against ZEN, apply indirect ELISA to study the performance parameter of the kit. RESULTS: The limited concentration of detection of the ELISA-kit was 1ng/ml, linear range was 1-200 ng/ml, the linear equation was Y = 0.99 - 0.40 x (R2 = 0.99). The inhibition concentration of 50% against ZEN was 16.3 ng/ml. The average recovery rate of spiked corn and wheat was 96.5% and 95.5%, respectively, the coefficient of variant was 13.2% and 10.9%, respectively. The kit can be stored at 4 degrees C over 6 months. The cross reaction rate with the other mycotoxins was less than 1%, and coefficient of variant within-laboratory and between-laboratory was less than 15% and less than 20%, respectively. Detecting the VICAM sample with ELISA method and HPLC method, the results were within the range of the sample, and there was no statistic difference between the two methods. CONCLUSION: This ELISA-kit was quick, sensitive, stable and specific and can be used to determine ZEN in cereals.