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采用自制的抗细交链格孢菌酮酸(TA)多克隆抗体,建立了以辣根过氧化物酶催化鲁米诺-过氧化氢为发光体系的间接竞争化学发光酶联免疫分析方法定量检测谷物中TA毒素。通过研究选择最佳工作参数:反应缓冲液pH 6.4~7.4、包被浓度2.0μg/mL、封闭液1.0%OVA、抗血清稀释80000倍、竞争反应时间30 min、二抗稀释度为8000倍、发光底物125μL/孔。其线性回归曲线方程为y=49.82-20.14x,R~2=0.9966,半抑制浓度IC_(50)为0.973 ng/mL,最低检出限为0.010 ng/mL,线性范围0.032~30.244 ng/mL,在面粉和燕麦中的平均加标回收率为82.4%~96.1%和81.5%~93.4%,批内变异系数与批间变异系数均小于12%,与液质方法比较准确度无显著差异;与其他几种食品中的常见真菌毒素无交叉反应。可快速、准确、灵敏地测定谷物中TA检测。
Indirect competitive chemiluminescence enzyme-linked immunosorbent assay (ELISA) was developed for the determination of luminol-hydroperoxide-based luminol system using horseradish peroxidase-conjugated polyclonal antibody against streptozotocin (TA) Detect TA toxins in grains. The optimal working parameters were selected by research: reaction buffer pH 6.4 ~ 7.4, coating concentration 2.0μg / mL, blocking solution 1.0% OVA, antiserum diluted 80000 times, competitive reaction time 30 min, secondary antibody dilution 8000 times, Luminescent Substrate 125 μL / well. The linear regression curve equation was y = 49.82-20.14x, R ~ 2 = 0.9966, the IC 50 of half-inhibitory concentration was 0.973 ng / mL, the minimum detectable limit was 0.010 ng / mL and the linear range was 0.032-30.244 ng / mL , The average spiked recoveries in flour and oats were 82.4% -96.1% and 81.5% -93.4%, respectively, and the coefficients of variation between batches and batches were all less than 12%. There was no significant difference between the two methods. No cross-reactivity with common mycotoxins in several other foods. Fast, accurate and sensitive determination of TA in cereals.