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.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
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)