建立固相酶联免疫吸附试验技术诊断社区糖尿病患者人巨细胞病毒活动性感染  

Establishment of IgM antibody capture ELISA and its application in detecting active infection of human cytomegalovirus in patients with diabetes mellitus

作　　者：张建英 徐冉行[2] 林伟良[3] 刘文虎[3] 凌志强 

机构地区：[1]宁波市眼科医院检验科,浙江宁波315040 [2]宁波市第六医院检验科,浙江宁波315040 [3]宁波市江东区明楼街道社区卫生服务中心内科,浙江宁波315040 [4]Shiga University of Medical Sciences, Japan Otsu 520-2192

出　　处：《中华医院感染学杂志》2011年第1期201-204,共4页Chinese Journal of Nosocomiology

基　　金：浙江省医药卫生科学研究基金项目(2005B156)

摘　　要：目的建立检测人巨细胞病毒(HCMV)抗原特异性IgM的抗体捕捉酶联免疫吸附试验(Capture-ELISA),并分析糖尿病患者HCMV活动性感染状态。方法利用抗人IgM(μ链特异性)抗体包被固相载体,作为捕捉抗体吸附待检血清中的IgM,经过洗涤除去血清中IgG及其他成分,再加入特异性抗原与捕捉到的相应IgM抗体结合,然后加入酶标抗体和底物显色进行测定;同时,应用建立的酶联免疫捕获法诊断糖尿病患者HCMV近期感染状态,并与常用的间接酶联免疫吸附试验(In-ELISA)及RT-PCR进行比较。结果 Capture-ELISA的特异性及敏感性均高于间接法,且不受RF因子的影响。结论 Capture-ELISA操作简单、快速,且具有较高的特异性和灵敏度,是检测IgM抗体理想的方法之一。OBJECTIVE To establish IgM antibody capture enzyme-linked immunosorbent assay （Capture-ELISA） and to detect active infection of human cytomegalovirus （HCMV） in patients with diabetes. METHODS Antihuman IgM with μ chain specificity was coated on microplates as capturing antibody for absorbing IgM in the specimens, then add specific antigen for integrating with captured IgM after washing for removal of IgG and other component in serums, finally detection by chromogenic reaction between a enzyme labeled antibody and substrate. Capture-ELISA was used to detect HCMV infection in patients with diabetes mellitus, and the results were compared with that indirect enzyme-linked immunosorbent assay （Indirect-ELISA） and RT-PCR. RESULTS The specificity and sensitivity of MAC-ELISA were shown to be significantly higher than that of indirect ELISA, and its results were not affected by RF. CONCLUSION Capture-ELISA is simple, rapid and shous higher specificity and sensitivity, which is one of best method for detecting IgM.

关 键 词：人巨细胞病毒 抗体捕捉酶联免疫吸附试验 间接酶联免疫吸附试验 

分 类 号：R446.1[医药卫生—诊断学]