An enzyme immunoassay for the quantitative measurement of estradiol in serum and plasma.
Estradiol (1,3,5(10)-estratriene-3,17_-diol; 17_-estradiol; E2) is a C18 steroid hormone with a phenolic Aring. This steroid hormone has a molecular weight of 272.4. It is the most potent natural Estrogen, produced mainly by the Graffian follicle of the female ovary and the placenta, and in smaller amounts by the adrenals, and the male testes (1,2,3). Estradiol (E2) is secreted into the blood stream where 98% of it circulates bound to sex hormone binding globulin (SHBG) and to a lesser extent to other serum proteins such as albumin. Only a small fraction circulates as free hormone or in the conjugated form (4,5). Estrogenic activity is affected via estradiol-receptor complexes which trigger the appropriate response at the nuclear level in the target sites. These sites include the follicles, uterus, breast, vagine, urethra, hypothalamus, pituitary and to a lesser extent the liver and skin.
In non-pregnant women with normal menstrual cycles, estradiol secretion follows a cyclic, biphasic pattern with the highest concentration found immediately prior to ovulation (6,7). The rising estradiol concentration is understood to exert a positive feedback influence at the level of the pituitary where it influences the secretion of the gonadotropins, follicle stimulating hormone (FSH), and luteinising hormone (LH), which are essential for folicular maturation and ovulation, respectively (8,9). Following ovulation, estradiol levels fall rapidly until the luteal cells become active resulting in a secondary gentle rise and plateau of estradiol in the luteal phase. During pregnancy, maternal serum Estradiol levels increase considerably, to well above the pre-ovulatory peak levels and high levels are sustained throughout pregnancy (10). Serum Estradiol measurements are a valuable index in evaluating a variety of menstrual dysfunctions such as precocious or delayed puberty in girls (11) and primary and secondary amenorrhea and menopause (12). Estradiol levels have been reported to be increased in patients with feminising syndromes (14), gynaecomastia (15) and testicular tumors (16). In cases of infertility, serum Estradiol measurements are useful for monitoring induction of ovulation following treatment with, for example, clomiphene citrate, LH-releasing hormone (LH-RH), or exogenous gonadotropins (17,18).
During ovarian hyperstimulation for in vitro fertislisation (IVF), serum estradiol concentrations are usually monitored daily for optimal timing of human chorionic gonadotropin (hCG) administration and oocyte collection (19).
The DRG Estradiol sensitive ELISA Kit is a solid phase enzyme-linked immunosorbent assay (ELISA), based on the principle of competitive binding. The microtiter wells are coated with a polyclonal antibody directed towards an antigenic site on the Estradiol molecule. Endogenous Estradiol of a patient sample competes with an Estradiol horseradish peroxidase conjugate for binding to the coated antibody. After incubation the unbound conjugate is washed off. The amount of bound peroxidase conjugate is reverse proportional to the concentration of Estradiol in the sample. After addition of the substrate solution, the intensity of colour developed is reverse proportional to the concentration of Estradiol in the patient sample.