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|Method & Line||Sample & Target||Product||Package Info|
|MicroELISA||Plasma,Serum||EIAgen 17α OH-Progesterone Kit||Tests per Package: 96|
|EIAgen||The EIAgen 17α-OH Progesterone Kit has been designed for the quantitative determination of 17α-OH Progesterone Concentration in Human Serum or Plasma.||Code: LI10040F1||Package: 1 Microplate|
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The EIAgen 17α-OH Progesterone Kit has been designed for the quantitative determination of 17α-OH Progesterone Concentration in Human Serum or Plasma.
Plasma/Serum concentrations of 17α-hydroxyprogesterone (17α-OHP) are valuable in the initial diagnosis of congenital adrenal hyperplasia (CAH) 1, 2. This common inborn error of metabolism is usually characterized by deficiency in the C21-hydroxylase enzyme system, and necessitates steroid replacement therapy. Adequacy of treatment has been monitored by determining circulating 17α-OHP concentrations3,4.
The incidence is roughly estimated to be 1 in 15,000 newborns and can reach as high as 1 in 1480 in native Alaskans. Early diagnosis is valuable to detect CAH in newborns afflicted with the disease, not clinically recognizable but which will lead to life threatening adrenal crisis in the neonatal period and to determine the cause of infants with ambiguous genitalia. Delayed diagnosis may also lead to further virilization in female children, acceleration of skeletal maturation and premature development of secondary sex characteristics in male children. Prompt treatment can save the life of infants and allow afflicted children to attain normal growth.
17P is a steroid produced in the adrenal cortex and the gonads. It is the immediate precursor to 11-desoxycortisol (CpS) which is converted to cortisol. Because CpS is produced by 21-hydroxylation of 17P, measurement of 17P is an indirect indicator of 21-hydroxylase activity. CAH occurs where there is a deficiency of this enzyme. The result is a decrease in the conversion of 17P to CpS which blocks the normal synthesis of cortisol. Due to the feed back mechanism, a decrease in cortisol causes an increase in ACTH secretion resulting in adrenal hyperplasia. As 17P is not being converted, increased concentrations of this steroid will be found.
17P concentration increases during pregnancy in the maternal and fetal blood. After birth, values decline rapidly to reach normal adult values in 2 to 7 days. Thus it is advisable not to collect samples before the 3rd day of life. Premature and sick term infants exhibit 2 to 3 fold 17P values with no CAH disorder. It is suggested that a different cut off be adopted to pre-term and sick infants. In this method, a sample containing 17-OH progesterone is dispensed into a microplate well. An enzyme labeled 17OH progesterone derivative and biotinylated anti-17OH-progesterone are than added. After a suitable incubation, the antibody fraction is separated from unbound enzyme reagent.
The employment of several serum references of known 17-OH Progesterone concentration permits construction of a graph of activity and concentration. From comparison to the dose response curve, an unknown specimen's activity can be correlated with 17-OH Progesterone concentration.
The essential reagents required for a enzyme immunoassay include antibody, enzyme-antigen conjugate and native antigen.
Upon mixing biotinylated antibody, enzyme-antigen conjugate and a serum containing the native antigen, a competition reaction results between the native antigen and the enzyme-antigen conjugate for a limited number of antibody binding sites.
A simultaneous reaction between the biotin attached to the antibody and the streptavidin immobilized on the microwell occurs. This effects the separation of the antibody bound fraction after decantation or aspiration.
The enzyme activity in the antibody bound fraction is inversely proportional to the native antigen concentration. By utilizing several different serum references of known antigen concentration, a dose response curve can be generated from which the antigen concentration of an unknown can be ascertained.
The kit contains reagents for 96 tests (code LI10040F1).
Bring reagents to room temperature before use.
Conjugate Conc. 11X
17α-OH Progesterone Biotin
Wash Buffer Conc. 50X
Number of tests