摘要
目的 建立人血清中肌钙蛋白的检测方法。方法 将倍比稀释后的待检血清 2μl滴加于硝酸纤维膜上的方格内 ,5 %牛血清白蛋白 50℃下 30min封闭硝酸纤维膜 ,然后将该膜浸于 1∶1 0 0稀释的辣根过氧化物酶 (HRP)标记的抗人心肌肌钙蛋白的单克隆抗体N1 5C ,N1 6D ,37℃反应 40min后 0 .0 35MPBS -T液振荡洗涤 3次 ,每次 5min ,其后将膜浸入显色液中显色后水洗终止反应。目测判读结果。结果 辣根过氧化物酶标记的N1 5C单克隆抗体检测心肌梗塞患者血清 ,患者血清最高稀释为 1∶8,N1 6D检测患者血清时 ,血清稀释度可达 1∶64 ,用 2株单抗检测时正常人血清均无斑点出现。结论 由抗人心肌肌钙蛋白的单克隆抗体N1
Objective Development of a immunoassay for cardiac-specific troponin T(cTnT) from sera Method Serum samples and controls(2?μl) were diluted with multiple proportions and were dropped into the pane on the surface of the cellulose mitrate membrane,and the membrane was blocked with 5% bovine serum albumin for 30 min at 50℃. Then the membrane was immersed into 1∶100 diluted horseradish peroxidase labeled N 15C and N 16D mouoclonal antibody(McAb)(1∶100 diluted with pH7.4 and 0.035Mol PBS-T buffer solution) for 40 min at 37℃.The membrane was washed three times, 5 min in a intervals. After this the membrane was immerged into the chromogenic substrate to react and stop the reaction with running water. Dots were judged by visual method.Result When horseradish peroxidase labeled McAb N 15C was used to capture the cTnT of the serum from the myocardiac infarction patients, the serum titer was 1∶8; When N 16D used, the serum titer was 1∶64.The dots did not appeare when the serum of healthy persons was tested with McAb N 15C and N16D.Conclusion The dot-ELISA assay with N 16D McAb can be put into the use of qualitative immunoassay of cTnT.
出处
《新乡医学院学报》
CAS
2002年第3期164-166,共3页
Journal of Xinxiang Medical University
基金
河南省科委科技攻关项目 (991 1 60 2 0 5 )
河南省教委科技攻关项目 (1 99932 0 0 1 8)
