褪黑激素ELISA方法的建立  

Development of an ELISA for Melatonin

作　　者：张勇[1] 汪兴生[2] 夏银[2] 杨利国[2] 程瑞禾[2] 尚平[3] 何良军[2] 

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070 [2]南京农业大学动物科技学院,江苏南京210095 [3]南京军区总医院,江苏南京210002

出　　处：《中国兽医学报》2001年第6期579-580,共2页Chinese Journal of Veterinary Science

基　　金：江苏省自然科学基金资助项目 ( BK970 95 )

摘　　要：将褪黑激素 (ML T)与牛血清白蛋白 (BSA)连结的复合物 (ML T∶ BSA=12∶ 1)免疫 5只雄性新西兰大耳兔 ,收集高效价血清 ,并提纯抗体。在 96孔微量反应板上 ,以 1∶ 10 0 0 0的抗体包被 ,将 ML T与酶标 ML T先后加入到板中竞争结合抗体 ,制定标准曲线 ,建立了 ML T酶联免疫测定法 (ML T- EL ISA )。此方法灵敏度为 17.31pg,板内变异系数 8.1% ,板间变异系数 15 .4%。用建立的此法测定夏至时考力代羊昼夜血样 ,测定结果跟国外报道的基本一致 ;用放射免疫法检测同一批血样 ,也获得类似结果 。An enzyme labeled immunosorbent assay(ELISA) for determination of melatonin in animal blood was described.The rabbit antibody empolyed was raised to the conjugate formed by coupling melatonin to bovine serum albumin with formaldehyde reaction,96-well microtiter plates were coated with anti-melatonin antibody(1∶ 10 000),and then were added with melatonin and enzyme labeled malatonin(1∶300),finally a standard curve was created.The sensitivity of the method was 17.31 pg/well.The variation coefficients of intra-and inter-plates were 8.1% and 15.4% respectively.The 24 h blood sample of Corridale sheep in the summer solstice was tested by the method,the results were equal to the foreign.The results were also proved by MLT-RIA,indicating the ELISA for melatonin is reliable.

关 键 词：褪黑激素 ELISA 羊 

分 类 号：S858.26[农业科学—临床兽医学] S852.1[农业科学—兽医学]