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当前位置: 首页 > 文章 > 甲基对硫磷的酶联免疫吸附分析(ELISA)研究 农业环境科学学报 2005,24 (01) 187-190
Position: Home > Articles > Study on ELISA of Parathion-methyl Journal of Agro-Environment Science 2005,24 (01) 187-190

甲基对硫磷的酶联免疫吸附分析(ELISA)研究

作  者：
韩丽君;贾明宏;钱传范;李成文;张冰
单  位：
军事医学科学院;中国农业大学理学院
关键词：
酶联免疫吸附测定法;甲基对硫磷;农药残留
摘  要：
根据甲基对硫磷的结构特征,从甲基对硫磷的甲氧基部位连接了两种间隔臂,合成了两种半抗原,并通过活性酯法和混合酸酐法分别制备了与不同的载体蛋白偶联的人工抗原。采用其中两种结合物进行动物免疫,得到了针对两种抗原的高效价抗体。在此基础上,建立了两种甲基对硫磷的酶联免疫吸附测定(ELISA)方法。结果表明,该方法检测限分别为2.0和6.1ng·mL-1。两种抗体对甲基对硫磷表现了很高的特异性,除了对硫磷以外,对结构相似的几种有机磷农药均无交叉反应。土壤和河水中的添加回收实验结果表明,采用ELISA方法得到的回收率及CV值都与GC方法基本吻合,且检测灵敏度比GC还高,符合农药残留分析的要求。
译  名：
Study on ELISA of Parathion-methyl
作  者：
HAN Li jun1, JIA Ming hong1, QIAN Chuan fan1, LI Cheng wen2, ZHANG Bing1 (1.College of Science, China Agricultural University, Beijing 100094, China; 2.Academy of Military Medical Sciences, Beijing 100850, China)
关键词：
ELISA; parathion-methyl; pesticide residue
摘  要：
In order to get high specificity antibody, two kinds of haptens with similar structure of parathion-methyl were synthesized. These two haptens were coupled with two different spacer arms (6-amino-hexoic acid and trans-4-amino-methyl-cyclohexyl acid) through methoxyphosphate group of parathion-methyl. The Mixed Anhydrous Method and the Active Ester Method were used to couple the hapten with different kinds of proteins. The protein concentration and the conjugation ratio of the conjugates were measured. Two of the conjugates (MP-1-AE-BSA and MP-2-MA-HAS) with high conjugation ratios (1∶37 and 1∶24) were chosen as immunogen and another one (MP-1-MA-KLH) was chosen as coating antigen. Through the immune program, both of the conjugates yieldedhigh specific and high affinity antibodies for parathion-methyl, the titers of the antiserum were 1∶30000 and 1∶40000, respectively. ELISA methods were developed successfully with both of the two kinds of antiserum. From the standardcurve, the linear range of parathion-methyl were 2～500 ng·mL-1 and 6～700 ng·mL-1, with the lowest detection limits (I20) of 2.0 and 6.1 ng·mL-1, respectively. The cross reaction tests indicated that most of the analogues and the related compounds didn't show cross reaction to the antibodies, except that parathion was able to inhibit the antibodies to parathion-methyl. After simple extraction and concentration, the fortified recoveries of parathion-methyl in soil and water were detected with ELISA and GC-FPD method, respectively. The results showed that the fortified recoveries and the CV were similar for thetwo methods, the lowest detection limit for ELISA were higher than GC method. The ELISA methods could be applied in the residue analysis of parathion-methyl.

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