一种稳定、灵敏且适用于ELISA的四甲基联苯胺底物  

A stable and sensitive tetramethyl benzidine substrate for ELISA

作　　者：单晶辉[1] 李宁[1] 关振宏[1] 段铭[1] 卢强[1] 刘红全[2] 孟锐奇[2] 张茂林[1] 

机构地区：[1]人兽共患病研究教育部重点实验室吉林大学人兽共患病研究所,吉林长春130062 [2]广西民族大学化学与生态工程学院,广西南宁530006

出　　处：《检验医学》2011年第4期260-262,共3页Laboratory Medicine

基　　金：吉林省科技发展计划重点项目(20080930);吉林大学农学部引进人才科研启动基金资助项目(4305050102C1);广西科学基金资助项目[桂科(自)0728057]

摘　　要：目的评价一种适用于酶联免疫吸附试验(ELISA)的即用型自制四甲基联苯胺(TMB)底物的特性。方法比较1-Step Turbo TMB底物、A/B两组分混合物底物和自制底物与不同稀释度酶标抗体的反应性及其对包被抗体的检测敏感性,对1-Step Turbo TMB底物和自制底物进行稳定性比较。结果 3种底物的吸光度(A)值均随着酶标抗体稀释度的增大而显著下降;当酶标抗体稀释度相同时,自制底物的A值显著高于另外2种底物(P<0.05);在直接ELISA条件下,自制底物对包被抗体的检测敏感性可达15.63 ng/mL;自制底物37℃贮存60 d后的A值仍高于1-Step Turbo TMB底物37℃贮存5 d的A值。结论该即用型自制TMB底物在检测敏感性及稳定性上优于1-Step Turbo TMB底物。Objective To evaluate a self-made ready to use tetramethyl benzidine（TMB） substrate for enzyme-linked immunosorbent assay（ELISA）.Methods 1-Step Turbo TMB substrate,A/B two-component TMB substrate and a self-made ready to use TMB substrate were applied.The reactivity against different dilution conjugated antibodies,the sensitivity for coated primary antibodies and the stabilities of 1-Step Turbo TMB substrate and self-made TMB substrate were compared,respectively.Results The result data showed that absorbance（A） values of the 3 substrates decreased greatly along with the dilution of the conjugated antibodies,and the A value of self-made substrate was significantly higher than those of other 2 kinds of substrates（P0.05）.The sensitivity for the self-made substrate with the coated antibodies by direct ELISA was 15.63 ng/mL,which was lower than those for the other 2 substrates.The A values of the self-made substrate after 60 d at 37 ℃ were even higher than that of 1-Step Turbo TMB substrate after 5 d under the same temperature.Conclusions The self-made TMB substrate is more sensitive and stable than 1-Step Turbo TMB substrate.

关 键 词：四甲基联苯胺底物 稳定 灵敏 酶联免疫吸附试验 

分 类 号：R446.61[医药卫生—诊断学]