腹泻性贝类毒素大田软海绵酸液态芯片检测方法的建立

Establishment of detection of okadaic acid in diarrlletic shellfish by indirect competitive liquid array immunoassay method

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。

A method for the detection of okadaic acid （OA） by liquid array was established. Applying the principle of indirect competitive immunoassay, coupled with fluorescent beads coated with OA-OVA as a carrier and using luminex Xmap 200TM liquid array sys- tem to detect the content of OA and the sensitivity of the method. The specificity of the method was conformed by the detection of OA, DTX-I, GYM, PTX2, SPX1, YTX, STX and MC-LR. Meanwhile, the average recovery of samples added OA was conformed. This method can be used for detecting the limit of OA in shellfish. The results showed that the concentration detection limit to OA was 0. 129μg/L, and the dynamic ranges were 0.2- 63 μg/L, which were higher than the sensitivity of corresponding conventional Idc ELISA. There was not the cross reaction with other toxins except for DTX-1. The average recovery of samples added OA was between 84.96% -90.35%. This method can be used for detecting the limit of OA in shellfish.