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利用间接竞争法免疫学原理,以自制的大田软海绵酸(okadaic acid,OA)与卵清白蛋白(ovalbumin,OVA)的偶联物OA-OVA包覆微球为载体,同时自制生物化抗OA单克隆抗体,利用液态芯片(Luminex Xmap 200TM)系统建立OA液态芯片定量检测方法;同时测定了方法的灵敏性和特异性,并对贝类模拟样本进行了测试。结果显示:液态芯片法对OA的检测线性范围为0.2～63μg/L,最低检测值为0.129μg/L,均优于酶联免疫吸附法;除与DTX-1的交叉反应率为54.2%外,与其他几种用于检测的贝类毒素没有任何交叉现象;在加标浓度为6.25～400μg/L时,回收率为84.96%～90.35%,变异系数为3.20%～6.59%。因此,该方法可满足海产品中贝类样品OA限量标准检测。
Using the indirect competitive immunological principle, the self-made OA-OVA-coated microspheres were used as carriers for the self-made bio-chemical OA (OA) and ovalbumin (OVA) Monoclonal antibody was used to establish the quantitative detection method of OA liquid chip by using Luminex Xmap 200TM system. Meanwhile, the sensitivity and specificity of the method were also tested, and the shellfish samples were tested. The results showed that the linearity of liquid chip method for OA was 0.2 ~ 63μg / L, the lowest value was 0.129μg / L, which was better than that of enzyme-linked immunosorbent assay. Except for the cross-reaction rate with DTX-1 was 54.2% , No cross phenomenon with other shellfish toxins tested. The recoveries ranged from 84.96% to 90.35% and the coefficients of variation ranged from 3.20% to 6.59% at spiked concentrations ranging from 6.25 to 400 μg / L. Therefore, this method can meet the seafood standard shellfish sample OA limit of detection.