人凝血因子Ⅸ∶Ag酶联免疫测定法的建立

Establishment of Enzyme linked Immunosorbent Assay of Human Factor Ⅸ Antigen

采用抗人凝血因子Ⅸ单克隆和多克隆两种抗体，建立了人凝血因子Ⅸ抗原（ＦⅨ∶Ａｇ）酶联免疫测定法（ＥＬＩＳＡ）。标准曲线相关系数为０．９９３（Ｐ＝０．００１，ｎ＝５），测定范围（６．２５～１００）Ｕ／Ｌ，最低检测量１．５Ｕ／Ｌ；批内及批间变异系数分别为７．２５％和４．７３％，回收率９９．７％；与人血浆中其它蛋白质未见交叉反应。测得２０人份正常人血浆中ＦⅨ∶Ａｇ为（１０２７±１７２）Ｕ／Ｌ，与ＦⅨ促凝活性（ＦⅨ∶Ｃ）测得值比较，差异无显著性（Ｐ＞０．５）。重症乙型血友病人血浆ＦⅨ∶Ａｇ测得值与临床诊断相符。

An enzyme linked immunosorbent assay(ELISA) for measuring human coagulation factor Ⅸ antigen (FⅨ∶Ag) has been established,using both monoclonal and polyclonal antibodies to FⅨ.The standard curve has a correlation coefficent r=0.993(n =5).This assay enabled the determination of FⅨ∶Ag in concentration from 6.25U/L to 100U/L.The lowest value of detection was 1.5U/L.Within run and between run coefficients of variation were 7.25% and 4.73% respectively.The recovery rate was 99.7%.There was no cross reaction with other proteins in human plasma.The mean concentrations of FⅨ∶Ag in plasma of 20 normal subjects was (1027±172)U/L.When the results were compared to those obtained by the determination of FⅨ coagulate activity,the difference between two groups wanst significant( P >0 5).The FⅨ∶Ag of the plasma in patient with severe hemophilia B were tallied with clinical diagnosis.