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∶2000,半数抑制浓度（IC50）为6.99 ng/mL；该方法的最低检测限达40 ng/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1.5-40 ng/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。
This experiment used carbodiimide get egg albumin synthesis of artificial antigens cou-pling neomycin,through square test to determine the best antigen packaged concentration and an-tibody dilution ratio,mass concentration logarithm of neomycin as abscissa,inhibition rate of anti-body as the ordinate drawing standard curve,established ELISA fast detection techniques of neo-mycin 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 was 1∶200,the best dilution multiple of antibodies was 1∶2 000,half inhibitory concentration（IC50 ） was 6.99 ng/mL;The minimum detection limit of the method was 40 ng/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 in 1.5 - 40 ng/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.
出处
《中国畜牧兽医》
CAS
北大核心
2016年第12期3163-3169,共7页
China Animal Husbandry & Veterinary Medicine
基金
CNCA检验检疫行业标准制定计划项目(2014B369)
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
import and export food animals
neomycin
enzyme-linked immunosorbent assay（ELISA）
detection method
