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Endocrine function tests
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Hormones Definition : Chemical Nature : ♦ Steroids
Hormones are chemical signals produced by specialized cells, secreted into the blood stream, And carried to a target tissue. Chemical Nature : ♦ Steroids Steroids hormones are lipid molecules that have cholesterol as a common precursor . - Steroids are produced by four organs: adrenal glands, ovaries, testes, and placenta.
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♦ Protein Protein hormones may be either peptides or glycoproteins . The peptides include insulin, glucagon, parathyroid hormone (PTH), growth hormone (GH), and prolactin (PRL). The glycoprotein hormones, follicle-stimulating hormone (FSH), luteinizing hormone (LH), thyroid-stimulating hormone (TSH), and human chorionic gonadotropin (hCG). ♦ Amines The amines are derived from amino acids and include epinephrine, norepinephrine, thyroxine, and triiodothyronine.
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Hormone tests methodology
Bioassays Before the development of modern assay methods, the detection and quantitation of hormones were based on expensive assays that depended on a biologic response from a live animal or tissue / organ culture. Competitive Protein Binding Competitive protein-binding (CPB) assays are based on competition for a limited number of protein-binding sites between a known added amount of labeled hormone and the unlabeled hormone present in the patients sample .
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Immunoradiometric Assay (IRMA)
Radioimmunoassay - RIA is a CPB technique that uses radio-labeled hormone and antibody prepared against the specific hormone as a binding site . - Competition between unlabeled hormone in the patient sample and the added-labeled hormone for a limited number of antibody-binding sites forms the basis of the assay . Immunoradiometric Assay (IRMA) - IRMA is similar to RIA in that a radiolabeled substance is used in an antibody-antigen reaction. However, the radioactive label is attached to the antibody instead of the hormone. - IRMA = sandwich technique form the “sandwich’’ comprising antibody- hormone –labeled antibody The amount of hormone present is directly proportional to the amount of radioactivity measured in the assay .
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Enzyme linked Immunosorbent Assays
- Enzyme linked immunosorbent assays (ELISA) have an enzyme label - Heterogeneous immunoassays ( Use a solid phase as the separation technique eg : micro titer well ) - Detection method based on enzyme catalyzed reaction then spectrophotometer is usually used to measure the color changes that are a result of enzyme activity . Enzyme-Multiplied Immunoassay Technique - Enzyme-Multiplied Immunoassay Technique (EMIT) have an enzyme label - Homogeneous assay (no physical separation of the bound and free components)
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