Position: Home > Articles > Establishment of ELISA Detection Method for Neomycin in Import and Export Animal
China Animal Husbandry & Veterinary Medicine
2016,43
(12)
3163-3169
进出口动物中新霉素ELISA检测方法的建立
作 者：
孙明君;邱芳;张金玲;郑小龙
单 位：
山东出入境检验检疫局检验检疫技术中心;潍坊出入境检验检疫局
关键词：
进出口食用动物;新霉素;酶联免疫吸附试验;检测方法
摘 要：
应用碳二亚胺法合成人工抗原得到卵清蛋白偶联新霉素,通过方阵试验确定最佳抗原包被浓度与抗体稀释倍数,以新霉素质量浓度对数为横坐标、抗体抑制率为纵坐标绘制标准曲线,建立了进出口食用动物中新霉素ELISA快速检测技术,并对建立的方法进行了回收率和特异性试验。结果表明,建立的检测方法包被人工合成抗原的最适稀释度为1∶200,抗体的最佳稀释度为1∶2 000,半数抑制浓度(IC50)为6.99ng/mL;该方法的最低检测限达40ng/mL,与庆大霉素、卡那霉素、链霉素、托普霉素、阿米卡星、双氢链霉素的交叉反应率均小于0.1%,平均加标回收率83.77%。标准曲线在1.5~40ng/mL范围内是线性的,相关系数为0.987,标准曲线方程为y=-37.66x+94.592;与常规液相检测方法相比操作简单、快速。本研究建立了一种快速、高效、特异的新霉素ELISA检测方法。
译 名：
Establishment of ELISA Detection Method for Neomycin in Import and Export Animal
作 者：
SUN Ming-jun;QIU Fang;ZHANG Jin-ling;ZHENG Xiao-long;Inspection and Quarantine Technology Center of Shandong Entry-exit Inspection and Quarantine Bureau;Weifang Entry-exit Inspection and Quarantine Bureau;
关键词：
import and export food animals;;neomycin;;enzyme-linked immunosorbent assay(ELISA);;detection method
摘 要：
This experiment used carbodiimide get egg albumin synthesis of artificial antigens coupling neomycin,through square test to determine the best antigen packaged concentration and antibody dilution ratio,mass concentration logarithm of neomycin as abscissa,inhibition rate of antibody as the ordinate drawing standard curve,established ELISA fast detection techniques of neomycin in edible animal for import and export.Methods of recovery and specific test had been done in further research.Results showed that the test dilution ratio of optimum synthetic antigen was1∶200,the best dilution multiple of antibodies was 1∶2 000,half inhibitory concentration(IC50)was 6.99ng/mL;The minimum detection limit of the method was 40ng/mL,and cross reaction rate of gentamicin,kanamycin,streptomycin,tobramycin,amikacin and dihydrostreptomycin were less than 0.1%,the average of recovery rate was 83.77%.The range of standarding curve in1.5~40ng/mL was linear,the correlation coefficient was 0.987,the equation of standarding curve was y=-37.66x+94.592;Compared with HPLC method,the operation was simple and fast.The test results showed that the enzyme-linked immunosorbent method of detecting neomycin was rapid,efficient and specific.
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