人绒毛膜促性腺激素游离β亚单位磁分离酶联免疫方法的建立  

Methodological study on magnetic enzymic immunoassay for detecting free hCGβ subunit

作　　者：刘振世[1,2] 周昊[3] 陈海生[3] 谢晶[3] 卢新[4] 杨祥良[4] 

机构地区：[1]华中科技大学生命科学与技术学院 [2]北京倍爱康生物技术股份有限公司,北京100070 [3]北京倍爱康生物技术股份有限公司 [4]北京海淀妇幼保健医院

出　　处：《细胞与分子免疫学杂志》2006年第3期392-394,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家科技攻关计划资助(No.2003BA10A05)

摘　　要：目的建立一种新的测定人绒毛膜促性腺激素游离β亚单位(hCGβ)磁分离酶联免疫分析法(MEIA)。方法用识别不同表位的两株单克隆抗体(mAb),一株mAb用FITC标记,另一株用碱性磷酸酶(AP)标记;以偶联羊抗FITC抗体的磁珠作为固相,以酚酞单磷酸酯溶液为底物,建立测定hCGβ磁分离酶免疫测定法。结果本MEIA法的灵敏度为0.1IU/L,批内CV小于8.5%,批间CV小于14%,平均稀释回收率为92.5%;与促黄体生成激素(leuteinizinghormone,LH)、促甲状腺激素(thyroidstimulatinghormone,TSH)和促卵泡成熟激素(folliclestimulatinghormone,FSH)均无交叉反应,与完整的hCG(1000IU/L)的交叉反应为1.2%,有效期不低于14个月。结论本方法的性能优于现有的放射免疫(RIA)和ELISA试剂盒,接近进口同类试剂的水平,有助于为市场提供一种高质量而价廉的hCGβ测定试剂盒。AIM： To establish a novel magnetic enzyme immunoassay （MEIA） for detecting human choriogonadotropin freeβ subun-it（hCGβ）. METHODS： Two monoclonal antibodies（mAb） were used for conjugating with FITC and with alkaline phosphatase（AP） respectively, which incorporated magnetic solid phase separation. Magnetic beads werecoupled with sheep anti-FITC antibody assolid phase, and phenolphthalein monophosphate was used as substrate to set up MEIAfor detecting hCGβ, RESULTS： The sensitivity of hCGβ MEIA kit reached 0.1 IU/L. Theintraassay variation and inter-assay variation were 8, 5% and 14% respectively, withthe average recovery rate of dilution of 92.5%. The kit showed no cross reactivity toLH, TSH and FSH, while to intact hCG at concentration of 1000 IU/L cross reactivity was 1.2%. The time of efficacy of hCGβ MEIA kitwas longer than 14 months. CONCLUSION： hCGβ MEIA kit is better than hCGβ radio immunoassay and ELISA kit, which can provide a quality and cheaper hCGβ kit for market.

关 键 词：人绒毛膜促性腺激素游离β亚单位 磁分离酶免疫测定法 碱性磷酸酶 

分 类 号：R392.11[医药卫生—免疫学]