高灵敏度玉米赤霉烯酮时间分辨荧光快速定量检测卡的研发和应用

Development and Application of High Sensitivity Zearalenone Time-resolved Fluorescence Rapid Quantitative Detection Card

成功研发了一种灵敏度高、准确度好、快速、简易的双T线的玉米赤霉烯酮(ZEN)时间分辨荧光快速定量检测卡。通过对比ZEN检测卡检测线数量(单T线、双T线)的灵敏度,主要利用双T线的增补作用优势抬高灵敏度。对比了独立C线、非独立C线体系工艺,利用非独立C线工艺,C值与T值呈现负相关来提高灵敏度。筛选出羊抗鼠-自身鼠抗非独立C线工艺,在其基础上进一步筛选标记抗体量(5、10、20、30μg/mL),选择标记抗体量10μg/mL效价25429,10~500μg/kg抑制率33.57%~98.30%,灵敏度、效价最佳。用液相色谱与玉米赤霉烯酮(ZEN)时间分辨荧光快速定量检测卡同时检测不同基质的样本,两者的符合度很高,准确度在90.89%~119.33%,浓度CV6.18%。研究表明,用荧光层析检测卡检测粮食谷物中玉米赤霉烯酮的分析方法稳定性好,货架期符合要求,可测范围宽,具有很好的应用前景。

A double T-wire time-resolved fluorescence rapid quantitative detection card of zearalenone with high sensitivity,good accuracy,fast and simple was successfully developed.By comparing the sensitivity of the number of detection lines(single T line,double T line)of ZEN detection card,the main advantage of double T line supplement is used to raise the sensitivity.The process of independent C-wire and non-independent C-wire system is compared.By using the non-independent C-wire process,the C value is negatively correlated with the T value to improve the sensitivity.The non-independent C-line process of sheep anti-mouse and automouse anti-mouse was screened,and the labeled antibody amount was further screened(5、10、20、30μg/mL),and the labeled antibody amount was 10μg/mL,the titer was 25429,and the inhibition rate of 10~500μg/kg was 33.57%~98.30%,and the sensitivity and titer were the best.HPLC and zellalenone(ZEN)time-resolved fluorescence rapid quantitative detection card were used to detect the samples of different substrates at the same time.The agreement between the two was high,the accuracy was 90.89%to 119.33%,and the concentration was CV6.18%.The results show that this method has good stability,good shelf life and wide measuring range for the determination of zalalenone in grain.