β-人绒毛膜促性腺激素光激化学发光免疫分析方法的建立

Development of light initiated chemiluminescent immunoassay for beta-human chorionic gonadotrophin

目的建立一种检测人血清β-人绒毛膜促性腺激素（β-HCG）浓度的光激化学发光免疫分析方法。方法采用2株针对不同抗原表位的抗β-HCG抗体,其中一株抗体用来包被发光微粒,另一株抗体标记生物素,以双抗体夹心法检测人血清中的β-HCG浓度,并与Beckman-Coulter公司试剂（化学发光法）进行比较。结果发光微粒浓度为100μg/mL、生物素标记抗体浓度为7μg/mL时,检测范围为0~1 000 mIU/mL,灵敏度为0.16 mIU/mL,平均回收率为100.3%,批内变异系数（CV）为0.80%~3.99%,批间CV为2.25%,与TSH、FSH和LH的交叉反应率低,稳定性较好,与化学发光法的符合率好（r=0.99）。结论光激化学发光免疫分析方法测定β-HCG具有较高灵敏度、精密度和准确性,与化学发光法的符合率较好,适用于临床测定。

Objective To develop a light initiated chemiluminescent immunoassay for human beta-human chorionic gonadotrophin（β-HCG）.Methods Two monoclonal antibodies with different epitopes to β-HCG were selected.One was coated on luminescent microspheres,and the other was biotinylated.The β-HCG concentration in human sera was determined by double antibody sandwich light initiated chemiluminescent immunoassay,and the results were compared with Beckman-Coulter reagent（chemiluminescent immunoassay）.Results The detection range of the assay was 0-1 000 mIU/mL,when the concentrations of the luminescent microspheres and the biotinylated antibody were 100 μg/mL and 7 μg/mL respectively.The sensitivity was 0.16 mIU/mL,and the recovery rate was 100.3%.The within-run and between-run coefficients of variation（CV） were 0.80%-3.99% and 2.25% respectively.The cross reaction rates of the assay with thyrotropic-stimulating hormone（TSH）,follicle-stimulating hormone（FSH） and luteotropic hormone（LH） were low.The stability of the detection method was good.The coincidence rate with the chemiluminescent immunoassay was highly significant（r=0.99）.Conclusions The light initiated chemiluminescent immunoassay for human β-HCG have good sensitivity,precision and accuracy.The coincidence rate with Beckman kits is good,and this assay can be applied for clinical determination.