Development of an immunoenzymometric assay (IEMA) for the estimation of human thyroid stimulating hormone (hTSH) in serum.
A sensitive immunoenzymometric assay (IEMA) of serum thyrotropin (hTSH) was developed using anti-hTSH rabbit polyclonal antibody and anti-hTSH in-house monoclonal antibody with a sensitivity of 0.12 mIU/L. Serum samples were incubated in ELISA wells precoated with polyclonal antibody. The hTSH bound to the wells was incubated with monoclonal antibody (detector antibody) and further with goat anti-mouse antibody-horse radish peroxidase (GAM-HRP), which obviates the need to label the detector antibody. The assay was validated by recovery, linearity, and cross-reactivity experiments with a working assay range of 0.15 to 100 mIU/L and <10% coefficient of variation (CV) for both intra- and interassay. Good correlations were obtained when compared with Immunotech hTSH IRMA (r = 0.971, n = 35). This in-house ELISA can be used as an initial screening test for thyroid dysfunction.
Department of Atomic Energy (DAE), Radioimmunoassay Group, Radiopharmaceuticals Programme, Board of Radiation and Isotope Technology, Navi Mumbai, India. shubhangi.mirapurkar@rediffmail.com
Shubhangi
S
Mirapurkar
G
G
Samuel
N
N
Sivaprasad
D000906:Antibodies; D003429:Cross Reactions / Q000276:immunology; D006735:Horseradish Peroxidase; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D015203:Reproducibility of Results; D013960:Thyroid Function Tests / Q000379:methods*; D013972:Thyrotropin / Q000097:blood*
D000906:Antibodies; D013972:Thyrotropin; D006735:Horseradish Peroxidase