【摘要】 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更多还原

【Abstract】 Objective:To establish the sensitive, specific, stable and convenient sCD40 ELISA kit and explore the clinical value for measurement of sCD40. Methods:The mAb 5C11 against human CD40 was used as coating antibody, and the other mAb 3G3 recognizing a different epitope was selected to be labeled by biotin. The established ELISA Kit was used to detect the levels of sCD40 in sera from normal volunteers, patients with hyperthyroidism, chronic nephritis, leukemia, rheumatoid arthritis and lung cancer, pleural fluid from patients with lung cancer as well as culture supernatant of multiple myeloma and lymphoma cell lines. Results:sCD40 ELISA kit was successfully established and its measuring threshold was 15.6 pg/ml. The sensitivity did not decline after the kit was placed at 4℃ for 3 months. The normal serum concentration of sCD40 in healthy donors was 105.00±18.88 pg/ml, whereas higher levels of sCD40 in patients with hyperthyroidism, chronic nephritis, leukemia, rheumatoid arthritis and lung cancer were detected. Conclusion:A specific and sensitive ELISA kit for detecting human sCD40 was produced, which showed a potential value for monitoring some disease related to abnormal CD40 expression of signaling.更多还原