结核分支杆菌特异性抗原重组蛋白MPT63检测系统的建立  

Establishment detection system of recombination protein MPT63 on mycobacterium tuberculosis specific antigen

作　　者：程国平[1] 李克生[2] 张润玲[3] 

机构地区：[1]河南科技大学第一附属医院检验科,洛阳471000 [2]甘肃省医学科学研究院生物技术中心 [3]原兰州大学第二附属医院检验科

出　　处：《中国实用医刊》2013年第2期27-29,共3页Chinese Journal of Practical Medicine

摘　　要：目的建立规范的结核分枝杆菌（MTB）特异性抗原重组蛋白MPT63酶联免疫吸附试验（EHSA）检测系统。方法采用方阵法确定蛋白包被浓度和酶标二抗浓度，以ROC曲线法确定MPT63蛋白EHSA法检测的cut一0ff值。探讨规范的EHSA检测系统建立的方法。结果EUSA检测系统M蹦3蛋白包被浓度1：128000，酶浓度1：1000时为最佳；在cut—ofr值为0．35—0．40时检测MPT63蛋白抗体的灵敏度和特异性最为理想，分别是75．0％和78．6％。结论用方阵法确定蛋白包被浓度和酶标二抗浓度，并以ROC曲线法确立cut—ofr值是建立规范化的EMSA检测系统的关键环节。Objective To set up a standard testing system of mycobacterium tuberculosis specif- ic antigen- recombinated MPT63 protein by enzyme-linked immunosorbent assay method. Methods The square matrix method was used to determine the concentration of coated protein and the concentration of enzyme labeling antibody. Receiver operating characteristic method was used to determine the cut-off val- ue of ELISA detected MPT63 protein. To approach an establishment of standard ELISA detection system. Results ELISA detected MPT63 protein may attain higher sensibility and specificity when the coated protein was diluted according to 1 ： 128 000 and the enzyme labeling antibody was diluted by 1 ： 1000. Conclusions The critical elements in ELISA detection system are the application of square matrix meth- od to determine the concentration of coated protein and the concentration of enzyme labeling antibody and application the ROC to determine cut-off.

关 键 词：重组蛋白MPT63 酶联免疫吸附试验 ROC曲线 cut—off值 

分 类 号：R4[医药卫生—临床医学]