Development of light initiated chemiluminescent immunoassay for beta-human chorionic gonadotrophin
2010
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.
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