Document: K217IE Instruction version: 411
Cat.# K217 17-OH-progesterone EIA Format version 307
Manufacturer: Xema-Medica Co. Ltd, P.O.Box 58, 105043 Moscow, Russia
Telephone/fax +7 (095) 152-3731, 152-3160; email email@example.com internet www.xema.ru
17-ÎÍ-progesterone (17-ÎÍ-P) is a steroid with molecular weight 330 D, an intermediate precursor in biosynthesis of glucocortiosteroids, estrogens and androgens.
17-OH-P is secreted by adrenals, ovaries and testicles by the enzyme 21-hydroxylase. 17-ÎÍ-Ð is secreted by ovaries during follicular phase; its serum level remains stable by the end of luteal phase. In case of non-fertile cycle, the serum level of 17-OH-P decreases; in case of fertilization, this hormone is secreted by corpus luteum.
The determination of 17-OH-P is important for diagnosis of inborn adrenal hyperplasia which causes elevated secretion of androgens and the development of adrenogenital syndrome (AGS). In AGS, the deficient 21-hydroxylase activity causes blocked steroid synthetic pathway and correspondent dramatic increase in serum 17-OH-P level. If the deficiency of 21-hydroxylase is acquired in mature age, or in case of delayed inborn defect, the serum 17-OH-P may remain normal.
This test is based on competition enzyme immunoassay principle. Tested specimen is placed into the microwells coated by specific anti-17-OH-progesterone-antibodies simultaneously with conjugated 17-OH-progesterone-peroxidase. 17-OH-progesterone from the specimen competes with the conjugated antigen for coating antibodies. After washing procedure, the remaining enzymatic activity bound to the microwell surface is detected and quantified by addition of chromogen-substrate mixture, stop solution and photometry at 450 nm. Optical density in the microwell is inversely related to the quantity of the measured analyte in the specimen.
Materials required but not provided
- Distilled or deionized water;
1. INFECTION HAZARD: There is no available test methods that can absolutely assure that Hepatitis B and C viruses, HIV-1/2, or other infectious agents are not present in the reagents of this kit. All human blood products, including patient samples, should be considered potentially infectious. Handling and disposal of this material should comply with the rules defined by an appropriate local biohazard safety guidelines.
2. Avoid contact with 5% H2SO4. It may cause skin irritation and burns.
3. Do not use reagents after expiration date.
4. Do not mix or use components from kits with different lot numbers.
5. Replace caps on reagents immediately. Do not swap caps.
6. Do not pipette reagents by mouth.
1. Store the whole kit at 2 to 8° C upon receipt until the expiration date.
2. After opening the pouch keep unused microtiter wells TIGHTLY SEALED BY ADHESIVE TAPE (INCLUDED) to minimize exposure to moisture.
Microplate photometer with 450 nm wavelength and OD measuring range 0-3.0.
Dry thermostate for 37Ñ
SPECIMEN COLLECTION AND STORAGE.
1. This kit is intended for use with serum or plasma (ACD- or heparinized). Grossly hemolytic, lipemic, or turbid samples should be avoided.
2. Specimens may be stored for up to 48 hours at 2-8° C before testing. For a longer storage, the specimens should be frozen at -20° C or lower. Repeated freezing/thawing should be avoided.
1. All reagents (including unsealed microstrips) should be allowed to reach room temperature (20 to 25oC) before use.
2. All reagents should be mixed by gentle inversion or vortexing prior to use. Avoid foam formation.
3. It is recommended to spin down shortly the tubes with calibrators on low speed centrifuge.
4. Prepare Washing solution from the concentrate by 10x dilution in distilled water. Diluted Washing solution is stable up to 2 weeks at 2-8° C.
It is recommended that pipetting of all calibrators and samples should be completed within 3 minutes.
Alternative units: 1 nmol/l = 3,05 ng/ml
See the example of calibration curve in Quality Control insert.
Control sample(s) should fit into the ranges shown in QC insert (see attached).
Based on data obtained by Xema-Medica, the following normal ranges are recommended (see below). However, GLP rules recommend that each laboratory should establish its own reference range.