鲀毒鱼类中河鲀毒素直接竞争抑制性酶联免疫吸附试验测定方法的研究

Determination of tetrodotoxin in puffer fishes using monoclonal antibody-based direct competitive inhibition enzyme-linked immunosorbent assay

为检测毒鱼类中的河毒素 ,建立了用抗河毒素单克隆抗体检测河样品中河毒素的直接竞争抑制性酶联免疫吸附试验 (ELISA)方法。结果表明 ,直接ELISA法较间接ELISA法灵敏 ,最低检出浓度为 0 1ng mL(每检测孔 0 0 1ng) ,线性范围为 5～ 5 0 0ng mL ,方法的加标回收率为73 0 %～ 118 0 %。本法与传统的小鼠生物试验相比 ,两种方法的测定结果之间在统计学上差异无显著性 (配对t检验 ,P >0 1) ,并具有良好的相关性 (r =0 94 4)。对来自江苏省东海和长江水域的河样品的毒力进行了测定 ,结果表明本方法简单、快速、灵敏 。

Direct competitive inhibition enzyme\|linked immunosorbent assay(ELISA) for determination for tetrodotoxin in puffer fish samples was developed using anti\|tetrodotoxin monoclonal antibodies. Results showed that direct ELISA was sensitive than indirect ELISA,the minimum detectable concentration of tetrodotoxin was 0 1 ng/mL (0 01 ng/assay),and the linear range of standard curve was between 5 and 500 ng/mL. The recoveries from tetrodotoxin spiked samples of the method were 73 0%～118 0%. Compared with the traditional mouse bioassay system,It showed no significant difference between the two methods(for paired\| t test, P >0 1),and good correlations for tetrodotoxin were obtained( r =0 944). In addition,the toxicity of some puffer fish samples from the East China Sea and the Yangzi River in Jiangsu Province were determined. The results implied that this was an simple,fast and sensitive method which met the need of detection of tetrodotoxin in puffer fish. Author's address: Ji Rong, Institute of Food Safety Control and Inspection, Ministry of Public Health, Beijing 100021,PRC.