摘要
采用间接竞争法的原理和液相芯片技术平台,选用黄曲霉毒素B1(AFB1)多克隆抗体对AFB1的定量检测方法进行了探索。通过优化偶联抗原浓度,确定AFB1多抗临界饱和浓度和抗原抗体最佳孵育时间,建立了AFB1液相芯片定量检测方法。以通用的IC50值作为灵敏度衡量标准,其灵敏度为1.33ng/mL,以IC10作为最低检测限衡量标准,其最低检测限为0.15ng/mL,线性方程为y=0.000 6x+0.000 8。应用该方法对脱脂牛奶和全脂牛奶中的AFB1进行添加回收率检测,在2.0~16.0ng/mL添加水平下,平均回收率均大于75%,相对标准偏差介于2.80%-4.69%(n=7)之间。经过实际样品的检测,证明该方法灵敏、稳定、快速、简便,适用于大量样本的检测。
The quantitative detection of Aflatoxin B1(AFB1) was investigated on the microsphere array platform and indirect competition by using the Anti-Aflatoxin B1.Through optimizing the concentration and saturated concentration of AFB1-BSA and Anti-Aflatoxin B1 as well as incubation time of antigen and antibody,the method of quantitative detection of AFB1 was established.As referring to the detection sensitivity standard IC50,results showed that the detection sensitivity was 1.33 ng/mL.Meanwhile,as referring to the limit of detection standard IC10,the detection limition was 0.15 ng/mL.The linear equation was y=0.000 6x+0.000 8.By applying this original research to test the imitating contaminated skim and full milk,the recovery rate of the two kinds of milk was all greater than 75% under the AFB1 added level of 2.0-16.0 ng/mL,and the relative standard deviation was between 2.80%-4.69%(n=7).Overall,the quantitative analysis of AFB1 using the microsphere array technology was sensitive,stable,rapid and simple.The method was applicable to mass sample testing.
出处
《农药学学报》
CAS
CSCD
北大核心
2011年第4期394-401,共8页
Chinese Journal of Pesticide Science
基金
中国质检总局资助项目(2009IK152)