摘要
目的:应用点免疫金渗滤法筛选适宜胶体金标记的抗人IgG(以下简称二抗),并运用多个评价指标进行比较和分析。方法:选择三个不同公司的二抗用胶体金标记,根据棋盘滴定法确定最佳标记条件;运用点免疫金渗滤法对三种二抗的胶体金标记后检测效能进行比较,采用包虫病人和健康对照血清,用包虫病特异性抗原检测包虫病患者血清中的特异性抗体,评价最优的二抗,并与酶联免疫吸附测定法进行比较。结果:A、B、C三种二抗标记的最佳标记条件为:p H均为8.5,加入量分别为38.4、24、19.2μg/ml,综合评价二抗B检测效能最优。将其与酶联免疫吸附测定法检测效能进行比较,两种方法检测结果的Kappa=0.895(P<0.05),吻合度较强。结论:应用点免疫金渗滤法,以及本文提出的评价指标,能够筛选出最佳二抗,对检测试剂盒中二抗的选择具有重要的参考价值。
Objective： To screen the appropriate anti-human Ig G（ secondary antibody） for conjugating with colloidal gold by dot immunogold filtration assay,and put forward the method of using multiple evaluation indicators for secondary antibody comparing and analyzing. Methods： Three different secondary antibodies A,B and C from three different companies were screened. Firstly the titration was used to determine the optimal reaction conditions. Then three secondary antibodies were conjugated to the colloidal gold,tested with a dot immunogold filtration assay for hydatidosis specific antibody detection. The optimal conjugated secondary antibody were compared with the standard indirect enzyme-linked immunosorbent assay. Results： The optimal p H of three secondary antibodies were 8. 5,the binding capacity were 38. 4,24 and 19. 2 μg / ml colloidal gold. According to the comprehensive evaluation,the diagnostic effects of secondary antibody B was better than others. Its diagnostic effects in dot immunogold filtration assay was compared with enzyme-linked immunosorbent assay. The Kappa was 0. 895（ P〈0. 05） and showed the two methods were in good agreement. Conclusion： The appropriate secondary antibody for conjugating with colloidal gold could be screened by dot immunogold filtration assay and the evaluation indicators which put forward by this study,the screening method would have an important reference value for all kinds of colloidal gold test kit to screen the suitable secondary antibody.
作者
房彬彬
齐新伟
王黎
冯晓辉
FANG Bin-Bin QI Xin-Wei WANG Li FENG Xiao-Hui(Clinical Medical Research Institute of the First Affiliated Hospital, Xinjiang Medical University, Urumqi 830054, Chin)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2016年第12期1790-1792,1796,共4页
Chinese Journal of Immunology
基金
新疆医科大学第一附属医院科研奖励基金(2011YFY16)
关键词
包虫病
筛选
胶体金标记
抗人IgG
Hydatidosis
Screen
Colloidal gold conjugate
Anti-human IgG
