RESUMO
A direct competitive enzyme-linked immunosorbent assay (ELISA) with monoclonal antibody (MAb) was diagnosed with progesterone (P) level of human serum. In high concentrations and large amounts of displacer effect, this monoclonal antibody (MAb) can retain biological activity so that it can be specially combined with progesterone. Under conditions of the existing displaced agent, monoclonal antibody 11F8(3)H5 can maintain high specificity and affinity and can specifically bind progesterone in serum. Progesterone ELISA standard curve was calculated according to the following formula: Logit(y) = -1.358Log(x) + 0.4961, r = 0.9944. The serum progesterone values obtained by this method correlated well with those obtained by chemiluminescent immunoassay (CLIA): the correlative equation was y = 0.7804x + 0.7600, r = 0.9126.
Assuntos
Anticorpos Monoclonais/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Progesterona/sangue , Animais , Afinidade de Anticorpos , Feminino , Humanos , Hibridomas/imunologia , Medições Luminescentes , Camundongos , Camundongos Endogâmicos BALB C , Progesterona/imunologia , Sensibilidade e EspecificidadeRESUMO
In recent years, various chemiluminescent clinical immunoassay kits have been widely applied to the detection of hormones. However, a kit for chloramphenicol (CAP) is often absent from most commercial product lists, even though it is important to control the levels of CAP residues in foodstuffs too. Therefore, we describe a simple, solid-phase chemiluminescence immunoassay (CLIA) for the measurement of CAP in foodstuffs. A rabbit anti-CAP IgG is passively adsorbed onto the walls of polypropylene plates. The labeled antigen is horseradish peroxidase (HRP) conjugate of CAP. Luminol solution is used as the substrate of HRP. The light yield is inversely proportional to the concentration of CAP. The method has a similar sensitivity (0.05 ng/ml), specificity, precision, and accuracy to a conventional enzyme immunoassay (EIA). The intra-assay and inter-assay CVs of ten samples were <8% and <20%, respectively, and the analytical recovery of the method was 87-100%. The experimental correlation coefficient of dilution was found to be 0.999 using milk supernatant as buffer. The detection limit for the method was 0.1-10 ng/ml, and it displayed good linearity.