微粒子化学发光免疫测定促甲状腺素敏感性方法的建立与应用

A novel microbeads-based chemiluminescence immunoassay for detecting human thyrotropin

目的 :建立一种低成本、高敏感性检测血清促甲状腺激素(TSH)的微粒子化学发光免疫分析 (CLEIA)方法。方法 :采用碱性磷酸酶标记的抗TSHβ亚单位特异性单克隆抗体 (mAb) ,与FITC偶联的抗TSHα亚单位的另一mAb配对 ,构成双抗体夹心免疫结合 ,用抗FITC多抗免疫磁珠做固相分离载体 ,用金刚烷胺CSPD为发光底物的非均相免疫分析法。结果 :方法灵敏度 4 μIU/L ,批内变异系数 (CV)平均为7.4 5 % ,批间CV平均为 10 .4 5 % ,回收率为 91.4 %～ 10 2 .4 %。将实际样品测定与ACS 180  系统检测的结果相比较有较好的相关性。结论 :微粒子化学发光定量测定TSH的成本低、灵敏度及特异性高和稳定性好 。

AIM: To establish a low cost and sensitive microbeads-based chemiluminescence immunoassay (CLEIA) for detecting human thyrotropin. METHODS: Thyrotropin in sera was captured by an alkaline phosphase-labeled mAb against TSH β subunit and a FITC-labeled mAb against TSH α subunit. Captured thyrotropin was then isolated with immuno-magnetic beads conjugated with anti-FITC antibody and then quantified by chemiluminescence using adamanatane amine as luminescent substrate. RESULTS: The sensitivity of this assay is 0.004 mIU/L. The intra-assay CV and the inter-assay CV of the assay were 7.45% and 10.45%, respectively. The recovery rate was 91.4%-102.4%. The examination result of the assay correlates well with that of ACS-180  system. CONCLUSION: The CLELA for detecting TSH is low-cost, sensitive, specific and stable, and therefore may have a promising prospect in clinical application.