可溶性gp130酶联检测试剂盒的研制及其运用

Preparation of sgp130 ELISA kit and its application

目的 :建立灵敏、特异、稳定和简便的人可溶性gp130 (sgp130 )酶标检测试剂盒。方法 :采用本室制备的鼠抗人gp130单抗T2作为包被单抗 ,另一株识别不同抗原位点的单抗T12经生物素 (biotin)标记后作为检测抗体 ,建立双单抗夹心的人sgp130酶标检测方法 ,并分别测定了 40例健康供血员、40例甲亢及 47例慢性肾炎患者血清中sgp130的含量。结果 :成功地研制了人sgp130酶联检测试剂盒 ,其灵敏度为 10ng/ml。该试剂盒 4℃放置 3个月 ,离散度 (CV) <± 7 6 %,回收率为95 %～ 111%,提示检测方法具有良好的灵敏度、稳定性和准确性。用该试剂盒测得的血清中sgp130含量的 95 %正常值范围为5 36 92～ 2 87 88(ng/ml) ,甲亢及慢性肾炎患者血清中sgp130的含量分别为 937 16± 2 17 5 9和 80 6 4 5± 138 4 7(ng/ml) ,明显高于正常对照者 (P <0 0 1)。结论 :建立的人sgp130酶标检测试剂盒 ,能够对血清中sgp130进行准确定量 ,可为临床gp130相关性疾病的辅助诊断、疗效估价和判断预后提供有价值的生物学参数 ,具有良好的运用前景。

Objective:To establish the sensitive,specific,stable and convenient sgp130 ELISA kit.Methods:The mAb T2 against human gp130 was used as coating antibody;the other mAb T12 recognized different epitope with T2 was labeled by biotin,then a ELISA kit for detecting sgp130 was set up.Results:sgp130 ELISA kit is successfully established and its sensitivity is 10 ng/ml.After the kit is placed in 4℃ for 3 months,the kit's CV is less than ±7.6% and the retrievable rate is 95%～111%.These indicate that it has highly sensitivity,stability and accuracy.The normal serum concentration of sgp130 in healthy donors is 536.92～287.88(ng/ml),but there is higher in patients with hyperthyroidism(937.16±217.5) and chronic nephritis(806.45±138.47).Significant difference is found comparing with normal control(P< 0.01). Conclusion:sgp130 ELISA kit may be of value in basic studies and contribute to gp130 related diseases' diagnosis,curative effects' evaluation and prognosis in clinic.