Time-resolved immunofluorometry of serum hTSH with enhanced sensitivity.
Sensitive TSH immunoassays offer a clear advance in discriminating the TSH concentrations in serum between hyperthyroid and euthyroid individuals; they have been proposed as the best single screening test for thyroid disorders. We have developed a highly sensitive serum TSH TRFIA based on DELFIA technology. Three monoclonal antibodies (McAbs) directed against different epitopes of the TSH molecule were involved in this assay, of which, one McAb was used to coat clear microwells, and the other two were biotinylated' for signal generation after being bound by the europium labeled streptavidin. The europium label captured on the well surface was quantified by a routine dissociation-enhancement procedure. The fluorescence intensity was directly proportional to the serum hTSH concentration. The assay required two steps and could be completed within 5 h. The analytical sensitivity reached 0.002 mIU/L with a sample volume of 100 microL, the function sensitivity was 0.017 mIU/L. Measurements by the present method correlated well with that obtained by the ACS-180 chemi-luminescence immunoassay (CLIA). The discrimination of hyperthyroid patients from clinically euthyroid patients by the present method was much better than that by the routine IRMA.
Wu
Feng-Bo
FB
Department of Isotopes, China Institute of Atomic Energy, Beijing. wufb@etang.com
Han
Shi-Quan
SQ
He
You-Feng
YF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000737:chemistry; D000918:Antibody Specificity; D001710:Biotin / Q000737:chemistry; D003429:Cross Reactions; D005063:Europium / Q000737:chemistry; D015200:Fluoroimmunoassay / Q000379:methods*; D006041:Goats; D006801:Humans; D006980:Hyperthyroidism / Q000097:blood / Q000175:diagnosis*; D007074:Immunoglobulin G / Q000737:chemistry; D007457:Iodine Radioisotopes / Q000737:chemistry; D008163:Luminescent Measurements; D051379:Mice; D004369:Pentetic Acid / Q000031:analogs & derivatives* / Q000737:chemistry; D012680:Sensitivity and Specificity; D019809:Streptavidin / Q000737:chemistry; D013972:Thyrotropin / Q000097:blood* / Q000276:immunology
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007457:Iodine Radioisotopes; C060068:diethylenetriaminetetraacetic acid; D005063:Europium; D001710:Biotin; D004369:Pentetic Acid; D013972:Thyrotropin; D019809:Streptavidin