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目的 :建立灵敏、特异和稳定的人可溶性CD4 0 (sCD4 0 )酶标检测试剂盒及探讨其检测的临床意义。方法 :采用该室制备的鼠抗人CD4 0单抗 5C11作为包被抗体 ,另一株识别不同抗原位点的单抗 3G3经生物素 (biotin)标记后作为检测抗体 ,建立双单抗夹心的人sCD4 0酶标检测方法和对各种质控参数进行优化。在此基础上测定了健康供血员、甲亢、慢性肾炎、白血病、类风湿性关节炎患者的血清、肺癌患者血清和胸水及多种多发性骨髓瘤 (MM)、淋巴瘤细胞株培养上清中sCD4 0的含量。结果 :成功地研制了人sCD4 0酶联检测试剂盒 ,其灵敏度为 15 6pg ml。该试剂盒 4℃放置 3个月 ,离散度 (CV) <± 7 9% ,回收率为 95 %～ 114 % ,提示检测方法具有良好的灵敏度、稳定性和准确性。用该试剂盒测得血清中sCD4 0含量的 95 %正常值范围 (x±s)为 10 5 0 0± 18 88pg ml,甲亢、慢性肾炎、白血病、类风湿患者的血清、肺癌患者血清和胸水中sCD4 0的含量明显高于正常对照组 (P <0 0 1)。结论 :成功地研制了检测人sCD4 0酶标检测试剂盒 ,对一些与CD4 0分子表达或信号异常的疾病患者外周血检测结果显示 ,该试剂盒在临床检测中具有潜在的应用价值
Objective: To establish a sensitive, specific and stable human soluble CD4 0 (sCD40) ELISA kit and to explore its clinical significance. Methods: The mouse monoclonal anti-human CD4 0 McAb 5C11 was used as the coating antibody. Another monoclonal antibody labeled with biotin labeled with different antigenic sites was used as the detection antibody to establish a dual-antibody sandwich Human sCD4 0 enzyme detection method and a variety of quality control parameters optimization. On the basis of this, the serums, pleural effusion and multiple myeloma (MM) in serum of healthy donors, hyperthyroidism, chronic nephritis, leukemia and rheumatoid arthritis, serum of patients with lung cancer and pleural fluid were measured in the culture supernatant of lymphoma cell lines sCD40 content. Results: The human sCD40 ELISA kit was successfully developed with a sensitivity of 15 6 pg ml. The kit was placed at 4 ° C for 3 months with a CV of <± 7 9% and a recovery of 95% ~ 114%, indicating that the assay has good sensitivity, stability and accuracy. The 95% normal range of serum sCD40 levels (x ± s) measured by this kit was 10500 ± 1888 pg ml, with serums from patients with hyperthyroidism, chronic nephritis, leukemia, rheumatoid and lung cancer The content of sCD40 in water was significantly higher than that in the normal control group (P <0.01). Conclusion: The kit for detection of human sCD40 was successfully developed. The results of peripheral blood tests on some patients with abnormal expression or abnormality of CD4 molecule showed that the kit has potential value in clinical detection