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    • 5. 发明申请
    • DETERMINATION OF THE PRESENCE OF AN ANTIGEN, PARTICULARLY HLA-B27 ANTIGEN, AT THE SURFACE OF A CELL, PARTICULARLY LYMPHOCYTES, BY CYTOTOXICITY TEST COMBINED WITH FLUORESCENT STAINING
    • 确定抗原,特别是HLA-B27抗原,细胞特异性淋巴细胞表面,通过与荧光染色相结合的细胞毒性测试
    • WO1995030152A1
    • 1995-11-09
    • PCT/CA1995000258
    • 1995-04-28
    • CENTRE DE RECHERCHE, INFLAMMATION, IMMUNOLOGIE ET RHUMATOLOGIEROY, RaynaldHEBERT, Jacques
    • CENTRE DE RECHERCHE, INFLAMMATION, IMMUNOLOGIE ET RHUMATOLOGIE
    • G01N33/569
    • G01N33/56977Y10S436/80Y10S436/821
    • The present invention relates to a method of determining the presence of an antigen at the surface of cells, particularly MHC or HLA antigens, by combining the sensitivity of a cytotoxicity test to the easy-to-perform fluorescence detection, particularly, by using propidium iodide. The claimed method may but does not necessarily involve the purification of lymphocytes from a blood sample. The erythrocytes contained in the test blood sample may be simply lysed after the immune reaction and complement action is completed. Moreover, the formation of the immune complexes, the complement fixation and reaction and the staining of the cells may take place at the same time in the test tube. This method is therefore very simple and very fast to execute. The facility of reading the positive cells is also an advantage to the claimed method over and above the methods using dyes like eosin or trypan blue, because of the direct visualization of colored cells as positive cells. Another advantage of the claimed method resides in its versatility of reading. Quantitation and identification of the positive cells is rendered feasible by fluorescence microscopy as well as by flow cytofluorometry. The claimed method has been validated in the diagnosis of ankylosing spondylitis (AS) and reactive arthritis (RA) by typing lymphocyte HLA-B27.
    • 本发明涉及通过将细胞毒性试验的灵敏度与易于进行的荧光检测结合起来,特别是通过使用碘化丙啶来确定细胞表面,特别是MHC或HLA抗原的抗原存在的方法 。 所要求保护的方法可以但不一定涉及从血液样品中纯化淋巴细胞。 测试血液样品中含有的红细胞可以在免疫反应和补体作用完成后简单地溶解。 此外,免疫复合物的形成,补体固定和反应以及细胞的染色可以在试管中同时进行。 因此,该方法非常简单,执行速度非常快。 由于将有色细胞作为阳性细胞的直接可视化,读取阳性细胞的方法对于使用诸如曙红或台盼蓝之类的染料的方法来说也是所要求保护的方法的优点。 所要求保护的方法的另一个优点在于其阅读的多功能性。 通过荧光显微镜以及流式细胞荧光测定,使阳性细胞的定量和鉴定变得可行。 所要求的方法已经通过分型淋巴细胞HLA-B27在强直性脊柱炎(AS)和反应性关节炎(RA)的诊断中得到验证。