用Hela细胞检测Bip抗体间接免疫荧光法的建立

Found the indirect immunofluorescence of detecting immunoglobulin binding protein antibody with Hela cells

目的建立抗免疫球蛋白结合蛋白(Bip)抗体检测的间接免疫荧光法(IIF),为探讨其在类风湿关节炎(RA)中的意义做出方法学准备。方法以Hela细胞为抗原基质固定于载片上,加入待测稀释血清温育,待抗原抗体特异结合后加入荧光素标记的抗人IgG与待测Bip抗体结合,最后置荧光显微镜下观察Hela细胞胞浆有无特异荧光出现。结果RA患者血清检测中可见到Hela细胞胞浆中出现特异性荧光,而健康献血员血清检测中未发现Hela细胞胞浆中的特异性荧光,实验所见荧光模式与文献报道相符。结论以Hela细胞为抗原基质的间接免疫荧光法能够对血清中Bip抗体进行检测。

Objective To found the indirect immunofluorescence（IIF） of detecting immunoglobulin binding protein （Bip） antibody with Hela cells as antigenic sources and to provide the means for exploring the diagnostic value of Bip antibody in patients with rheumatoid arthritis （RA）. Methods Put the Hela cells onto slides and fix them,then add serum samples to incubate. Add in anti-human IgG antibody with dihydroxyfluorane when the Bip antigen and the Bip antibody have bound,after the second antibody and Bip antibody bound,to observe the specific fluorescence in the cytoplasm of Hela cells using fluorescence microscope. Results The specific fluorescence in the cytoplasm of Hela cells could be observed in patients＇ serum samples with RA and didn＇t appear in healthy persons＇. The specific fluorescence in this experiment was the same as it in other literature. Conclusion The indirect immunofluorescence（IIF） with Hela cells as antigenic sources can detect immunoglobulin binding protein （Bip） antibody in serum samples.