17a-Hydroxyprogesterone Kit CLIA

N° du produit ABIN504755

Ce kit ELISA Chemiluminescent est destiné à la mesure quantitative de Humain 17a-Hydroxyprogesterone.

Aperçu rapide pour 17a-Hydroxyprogesterone Kit CLIA (ABIN504755)

Antigène: 17a-Hydroxyprogesterone

Reactivité: Humain

Méthode de détection: Chemiluminescent

Type de méthode: Competition ELISA

Application: ELISA

Détail du produit 17a-Hydroxyprogesterone Kit CLIA

Fonction: Competitive Enzyme Immunoassay (TYPE 7): 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.

Analytical Method: Quantitative

Attributs du produit: The Quantitative Determination of 17-OH Progesterone Concentration in Human Serum or Plasma by a Microplate Chemiluminescence Immunoassay

Information d'application

Indications d'application: All products that contain human serum have been found to be non-reactive for Hepatitis B Surface Antigen, HIV 1&2 and HCV Antibodies by FDA required tests. Since no known test can offer complete assurance that infectious agents are absent, all human serum products should be handled as potentially hazardous and capable of transmitting disease. Good laboratory procedures for handling blood products can be found in the Center for Disease Control / National Institute of Health, "Biosafety in Microbiological and Biomedical Laboratories," 2nd Edition, 1988, HHS Publication No. (CDC) 88-8395.

Plaque: Pre-coated

Protocole:

Specimien Collection and Preparation:

The specimens shall be blood, serum or heparanised plasma in type and taken with the usual precautions in the collection of venipuncture samples. For accurate comparison to establish normal values, a fasting morning serum sample should be obtained. The blood should be collected in a redtop (with or without gel additives) venipuncture tube or for plasma use evacuated tube(s) containing heparin. Allow the blood to clot for serum samples. Centrifuge the specimen to separate the serum or plasma from the cells. Samples may be refrigerated at 2_x001E_8oC for a maximum period of five (5) days. If the specimen(s) cannot be assayed within this time, the sample(s) may be stored at temperatures of _x001E_20oC for up to 30 days. Avoid repetitive freezing and thawing. When assayed in duplicate, 0.050ml of the specimen is required.

Reagent Preparation:

1. Working Tracer Reagent - Stable for 1 year. Measure 0.7 ml of 17-OH Progesterone Tracer Reagent and add to the vial containing Steroid Tracer Buffer. Store at 2-8C. 2. Wash Buffer Dilute contents of wash solution to 1000ml with distilled or deionized water in a suitable storage container. Diluted buffer can be stored at room temperature (20-27(C) for up to 60 days. 3. Working Signal Reagent Solution - Store at 2 - 8(C. Determine the amount of reagent needed and prepare by mixing equal portions of Signal Reagent A and Signal Reagent B in a clean container. For example, add 1 ml of A and 1ml of B per two (2) eight well strips (A slight excess of solution is made). Discard the unused portion if not used within 36 hours after mixing. If complete utilization of the reagents is anticipated, within the above time constraint, pour the contents of Signal Reagent B into Signal Reagent A and label accordingly.

Test Procedure:

Before proceeding with the assay, bring all reagents, serum references and controls to room temperature (20 - 27(C). 1. Format the microplates wells for each serum reference, control and patient specimen to be assayed in duplicate. Replace any unused microwell strips back into the aluminum bag, seal and store at 2-8(C. 2. Pipette 0.025 ml (25 L) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0.050 ml (50l) of working Tracer Reagent to all wells. 4. Swirl the microplate gently for 20-30 seconds to mix. 5. Add 0.050 ml (50l) of the 17a-OH Progesterone Biotin Reagent to all wells. 6. Swirl the microplate gently for 20-30 seconds to mix 7. Cover and incubate for 45minutes at room temperature. 8. Discard the contents of the microplate by decantation or aspiration. If decanting, blot the plate dry with absorbent paper. 9. Add 350l of wash buffer (see Reagent Preparation Section), decant (tap and blot) or aspirate. Repeat four (4) additional times for a total of five (5) washes. An automatic or manual plate washer can be used. Follow the manufacturers instruction for proper usage. If a squeeze bottle is employed, fill each well by depressing the container (avoiding air bubbles) to dispense the wash. Decant the wash and repeat four (4) additional times. 10. Add 0.100 ml (100l) of working signal reagent solution to all wells (see Reagent Preparation Section). Always add reagents in the same order to minimize reaction time differences between wells. 11. Incubate at room temperature for five (5) minutes in the dark. 12. Read the relative light units in each well with a chemiluminescence microplate reader for 0.5-1.0 seconds. The results should be read within 30 minutes after adding the working Signal Reagent. Note: Dilute the samples suspected of concentrations higher than 20ng/ml 1:1 and 1:5 with 17-OH Progesterone 0 ng/ml calibrator or male patient serum pools with a known low value for 17-OH Progesterone.

Restrictions: For Research Use only

Stockage

Détail du antigène

Antigène: 17a-Hydroxyprogesterone

Autre désignation: 17a-hydroxyprogesterone (17a-OHP)

Classe de substances: Hormone

Sujet: Plasma/Serum concentrations of 17a-hydroxyprogesterone (17a-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 17a-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 con_x001F_centration. From comparison to the dose response curve, an unknown specimen's activity can be correlated with 17-OH Progesterone concentration.