ABSTRACT
An enzyme immunoassay kit based on monoclonal antibodies (Mab) has been developed for measuring thyroid-stimulating hormone in human serum and plasma. Laboratory trials demonstrated its high sensitivity (3.1 microIU/ml), specificity, reliability, and accuracy. The analysis is simple and rapid (2.5 h), which is convenient for clinical use. The kit permits differentiation between normal subjects and patients with primary hypo- or hyperthyroidism and helps monitor the treatment efficacy. Comparison with the Amerlite TSH-60 kit, carried out on 357 sera, showed a high correlation coefficient (0.96) between the two kits.
Subject(s)
Immunoenzyme Techniques , Reagent Kits, Diagnostic , Thyrotropin/blood , Antibodies, Monoclonal , Diagnosis, Differential , Humans , Hyperthyroidism/diagnosis , Hypothyroidism/diagnosis , Sensitivity and SpecificityABSTRACT
The solid-phase enzyme immunoassay for testosterone (TS), permitting the determination of this hormone at concentrations of up to 0.5 ng/ml, has been developed. The method comprises the adsorption of TS conjugated with soya trypsin inhibitor in the wells of a standard polystyrene assay plate, competition between adsorbed TS and TS under test for the binding sites of specific antibodies, and the detection of antibodies bound to the carrier by means of peroxidase-labeled antispecific antibodies. Antisera to TS have been obtained by the immunization of rabbits with TS conjugated with bovine serum albumin of a known composition. These antisera are specific to TS and do not interact with estrogens and progesterone. The study of their cross reactions with eleven TS derivatives has demonstrated that antibodies reveal the presence of structural changes in ring D of the molecule of TS and are insensitive to variations in ring A. The determinant comprising the 17-OH-group essentially contributes to the binding of antibodies.
Subject(s)
Haptens/analysis , Immunoenzyme Techniques , Testosterone/blood , Adsorption , Animals , Antibody Affinity , Antibody Specificity , Binding Sites, Antibody , Binding, Competitive , Calibration , Cross Reactions , Immune Sera/isolation & purification , Immunization/methods , Immunoenzyme Techniques/instrumentation , Immunoglobulin G/immunology , Rabbits , Species SpecificitySubject(s)
Antigens/classification , Animals , Antigens/analysis , Antigens/genetics , Forensic Medicine , Humans , Species SpecificityABSTRACT
The use of the indirect ELISA techniques did not ensure the sharp differentiation of the antigens of the blood groups A and B on the polystyrene sorbent by means of heteroimmune sera, though such differentiation could be achieved by means of monoclonal antibodies. The test system known as "the lectin-antibody sandwich" was found to have the optimum sensitivity and specificity permitting the detection of soluble ABH antigens. This variant of ELISA permitted the detection of blood group A antigen both in native biological materials and in traces of blood and saliva, thus making it possible to carry out its quantitative determination.
