河豚毒素单抗ELISA检测试剂盒的研制

宫慧芝; 计融; 江涛; 杨军

doi:10.11847/zgggws2005-21-12-08

河豚毒素单抗ELISA检测试剂盒的研制

Development of ELISA-kit of quantitative analysis for tetrodotoxin

摘要

摘要:
目的研制快速检测河豚鱼类中的河豚毒素(tetrodotoxin, TTX)试剂盒。
方法在生产的抗TTX单克隆抗体基础上, 利用间接竞争酶联免疫吸附方法研制TTX快速检测试剂盒。
结果该试剂盒的最低检出浓度为5ng/ml, 标准曲线的线性范围为5~500 ng/ml, 回归方程为Y=-0.3546x+1.1006, r=0.99(P < 0.05);河豚鱼25和100 ng/ml 2个水平的加标回收率为72.45%~111.24%;试剂盒常温下可保存225 d; 与牛血清白蛋白(BSA)无交叉反应; 实验室内的变异系数和实验室间的变异系数均 < 20%。
结论该试剂盒快速、简单、灵敏, 可满足实际工作需要。

Abstract:
Objective To develop a rapid, sensitive, quantitative ELISA-kit for detection of tetrodotoxin in pufferfish.
Methods It was determined by indirect competitive inhibition enzyme-linked immunosorbant assay on the base of monoclonal antibodies against tetrodotoxin.
Results The limited concentration of detection of the EL ISA-kit was 5 ng/ml, linear range was 5~500 ng/ml, the linear equation was Y=-0.3546x+1.1006, r=0.99(P < 0.05).The recovery rate of spiked pufferfish on the level of 25 ng/ml and 100 ng/ml was among 72.45%~111.24%.The kit could be stored at normal temperature in 225 days.It was no cross reaction with bovine serum albumin(BSA); and coefficient of variant within-laboratory and between-laboratory was less than 20%.
Conclusion The ELISA-kit was simple, fast, sensitive and could meet the demand of practical work.

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