3种酶联免疫分析法在蓖麻毒素定量测定中的比较  

Comparison of Three Enzyme-Linked Immunosorbent Assay Methods for Quantitative Determination of Ricin

作　　者：马小溪[1] 刘合珠[1] 唐吉军[1] 郭磊[1] 谢剑炜[1] 

机构地区：[1]军事医学科学院毒物药物研究所,北京100850

出　　处：《分析化学》2011年第5期685-689,共5页Chinese Journal of Analytical Chemistry

基　　金：国家重大新药创制科技专项"综合性新药研发技术大平台"课题(No.2009ZX09301-002)资助

摘　　要：建立了蓖麻毒素的3种酶联免疫定量分析法,即光吸收、荧光和化学发光免疫分析法,并系统优化了各项实验条件。结果显示:对于化学发光检测法,当酶标抗体HRP-4C13稀释倍数为1∶8000时可获得最佳的信噪比,且酶与底物反应3 min后信号趋于稳定。随后在各自优化的条件下将3种方法用于毒素的检测。比较结果表明:化学发光酶联免疫分析法除具有线性范围宽(0.02～5.5μg/L,r2=0.999)、灵敏度高(检出限为5 ng/L)的特点外,还具有简单、快速、体系稳定性好等优点。将本方法用于不同实际样品基质,如饮用水、碳酸饮料、奶粉、咖啡和血清中添加的蓖麻毒素的检测,其检出限为0.005～0.08μg/L,回收率为89.6%～108.8%,适于污染及中毒样品中痕量蓖麻毒素的定量分析。Three determination methods in enzyme-linked immunosorbent assay,namely optical absorption,fluorescent and chemiluminescent immunoassay,were established for quantitation of biotoxins protein ricin in various matrices.The detection conditions were systematically optimized.The results showed the best signal-to-noise（SNR） could be obtained for CLIA when dilution factor was 1 ∶ 8000;and the chemiluminescence signal was stable at around 3 minutes after horseradish peroxidase reacted with its substrate.Whereafter,the three methods were compared for determination of ricin under individually optimized conditions.The comparative results indicated that chemiluminescent immunoassay could provide wider linear range（from 0.02 μg/L to 5.5 μg/L with correlation coefficient of 0.999）,higher sensitivity（limit of detection was 0.005 μg/L）,and could be adopted as a simple,rapid and robust approach.The CLIA method was applied to measure the spiked ricin in water,carbonated beverage,milk powder,coffee and human serum samples.The limits of detection and the recoveries were from 0.005 to 0.08 μg/L and from 89.6% to 108.8%,respectively.The method was quite suitable for quantitative determination of trace amounts of ricin in contaminated or poisoned samples.

关 键 词：蓖麻毒素 酶联免疫分析 化学发光 

分 类 号：Q946[生物学—植物学]