改良碱性磷酸酶-抗碱性磷酸酶桥联酶染色微板法检测细胞表面抗原  

Improved APAAP Immunocytochemical Staining -Trace Cytotoxicity Test Plate Method for Detection of Cell Surface Antigens

作　　者：蒋红梅[1] 蔡美英[1] 

机构地区：[1]华西医科大学基础医学院免疫学教研室,成都610041

出　　处：《华西医科大学学报》2001年第1期143-144,147,共3页Journal of West China University of Medical Sciences

摘　　要：目的　对碱性磷酸酶 -抗碱性磷酸酶 ( APAAP)桥联酶染色常规玻片法进行改良 ,用于检测细胞表面抗原。方法　用微量细胞毒试验反应板 (简称微板 )代替载玻片作细胞载体进行 APAAP桥联酶染色 ,检测人外周血淋巴细胞表面 CD3、CD4、CD8、CD11a、CD11b、CD18和 CD5 4的表达。结果　微板法与玻片法检测结果无显著性差异 ( P均 >0 .0 5 ) ,有良好重复性 ,并将检测 7项指标所需的血量由 10 m l减至 2 m l,各种试剂 (一抗、二抗、APAAP复合物和底物 )用量由 5 0μl减至 5μl。结论　微板法节约了试剂及用血量 ,操作方便 ,效果良好 。Objective To improve the conventional slide metho d of alkaline phosphatase*anti alkaline phosphatase (APAAP) immunocytochemical staining for detection of cell surface antigens. Methods Trac e cytotoxicity test plates were utilized in the detection of CD3,CD4,CD8,CD11a, CD11b,CD18 and CD54 on lymphocytes by APAAP immunocytochemical staining. The per centages of positive cells were counted. Meanwhile, slides were used in the sam e objects and procedure. Results The results of trace test plate method were similar to those of slide method ( P >0.05, respectively). Th e re producibility was very excellent. The plate method decreased the volume of blood from 10 ml to 2 ml to detect 7 norms, and each reagent from 50μl to 5μl. Conclusion Trace test plate method may not only attain the result si milar to that of the slide method, but also save blood and reagents; it is easy to do, so it is more suitable for clinical examination and scientific research in hospitals or institutions at the basic level.

关 键 词：微量细胞毒试验反应板 酶免疫细胞化学技术 细胞表面抗原 APAAP 

分 类 号：R446.61[医药卫生—诊断学]