Progesterone and estro
gen are hormones that are produce in the ovary.
They are important in the regulation of menstrual cycle, maintenance of pregnancy, development of secondary sexual characteristics and other.
Thus checking their levels in our body is essential for us to determine if there are abnormalities present.
These two hormones can be detected using different methods and even different kinds of specimen. We could determine their level in the circulation using blood and even in the urine.
Using blood specimens we could perform different immunoassay such as Radio Immuno Assay (RIA) and Enzyme-linked Immunosorbent Assay (ELISA)........
Progesterone
Since progesterone helps in the regulation of the menstrual cycle we could check for progesterone level so as to determine if the patient has ovulated or not. Thus this test can be used to determine infertility of women.
Progesterone in pregnant women rises 10 to 20 times higher than the luteal phase peak. Thus progesterone test is used to determine the ovulation and detect problems in luteal phase. It can also be used to monitor progesterone therapy.
Tests...
Zimmermann reaction
In this procedure an expanded array of steroids can be determined as long as it has oxygen containing carbon-20 and a group on carbon 21. These steroids can be converted into 17-ketosteroid which is present in urine.
This technique involves hydrolysis of urine into acid and extraction using ethylene dichloride. After extraction it is dried and the residue is then reacted with zimmerman reagent (m-dinitrobenzene). It then forms a red colored solution and measured spectrophotmetrically at 520nm
Immunoassay technique
ELISA Progesterone test
The progesterone test uses ELISA to determine the progesterone level in serum. Using the principle of competitive binding the patient's progesterone and the standard progesterone competes with limited binding sites.
The excess progesterone which are not bound with the antibodies are then washed. After washing color reagent is then added to form colored solution and then incubated. After incubation stop solution is added and then measured spectrophotometrically at 450 nm.
Estrogen
Estradiol
Since estradiol is the true female hormone it is the only hormone essential to evaluate ovarian function.
Estradiol determination is associated with differentiation of the cause of amenorrhea. High levels are seen in patients with hyperfunction of the ovary.
Estriol
The major hormone present in pregnant women is estriol, thus determination of estriol level of pregnant women helps in monitoring dangerous pregnancy.
Genetic disorders are also associated with low levels of estriol, such as down syndrome, adrenal abnormalities and other trisomies.
Estrone
In menopause women the ovaries ceases the production of estradiol and in turn produces estrone. Thus non menopausal women have small amount of estrone in their circulation. Abnormal results may be associated with tumor and hypopituitarism.
Tests...
Kober reaction
Due to the increase estrogen specifically estriol in pregnancy urine is used to monitor the development of the fetus during pregnancy. The assay used in determining urine estrogen is the kober reaction.
This method involves the heating of a urine sample in a strong aqueous solution containing hydroquinone. After cooling and dilution the reddish brown solution is then measured for its absorbance.
Other color reactions ...
click here
Note:
Use of ampicillin or neomycin causes a decrease of urine estrogen because of the diminished hydrolysis of estriol conjugated. This is caused by the antibiotic that reduces the bacterial level in the urine.
Meprobamate, L-dopa, and phenolphthalein gives falsely elevated levels of estrogen in urine.
Immunoassay techniques
ELISA estradiol test
Measurement of estradiol in serum makes use of ELISA just like the progesterone tests. It employs the principle of competitive binding where the patient's estradiol competes with the standard control.
Then extra unbound estradiol is washed and then TMB is added to make a color blue solution. It is then incubated and then stop solution is added and then measured spectrophotometrically in 450 nm.
gen are hormones that are produce in the ovary.They are important in the regulation of menstrual cycle, maintenance of pregnancy, development of secondary sexual characteristics and other.
Thus checking their levels in our body is essential for us to determine if there are abnormalities present.
These two hormones can be detected using different methods and even different kinds of specimen. We could determine their level in the circulation using blood and even in the urine.
Using blood specimens we could perform different immunoassay such as Radio Immuno Assay (RIA) and Enzyme-linked Immunosorbent Assay (ELISA)........
Progesterone
Since progesterone helps in the regulation of the menstrual cycle we could check for progesterone level so as to determine if the patient has ovulated or not. Thus this test can be used to determine infertility of women.
Progesterone in pregnant women rises 10 to 20 times higher than the luteal phase peak. Thus progesterone test is used to determine the ovulation and detect problems in luteal phase. It can also be used to monitor progesterone therapy.
Tests...
Zimmermann reaction
In this procedure an expanded array of steroids can be determined as long as it has oxygen containing carbon-20 and a group on carbon 21. These steroids can be converted into 17-ketosteroid which is present in urine.
This technique involves hydrolysis of urine into acid and extraction using ethylene dichloride. After extraction it is dried and the residue is then reacted with zimmerman reagent (m-dinitrobenzene). It then forms a red colored solution and measured spectrophotmetrically at 520nm
Immunoassay technique
ELISA Progesterone test
The progesterone test uses ELISA to determine the progesterone level in serum. Using the principle of competitive binding the patient's progesterone and the standard progesterone competes with limited binding sites.
The excess progesterone which are not bound with the antibodies are then washed. After washing color reagent is then added to form colored solution and then incubated. After incubation stop solution is added and then measured spectrophotometrically at 450 nm.
Estrogen
EstradiolSince estradiol is the true female hormone it is the only hormone essential to evaluate ovarian function.
Estradiol determination is associated with differentiation of the cause of amenorrhea. High levels are seen in patients with hyperfunction of the ovary.
Estriol
The major hormone present in pregnant women is estriol, thus determination of estriol level of pregnant women helps in monitoring dangerous pregnancy.
Genetic disorders are also associated with low levels of estriol, such as down syndrome, adrenal abnormalities and other trisomies.
Estrone
In menopause women the ovaries ceases the production of estradiol and in turn produces estrone. Thus non menopausal women have small amount of estrone in their circulation. Abnormal results may be associated with tumor and hypopituitarism.
Tests...
Kober reaction
Due to the increase estrogen specifically estriol in pregnancy urine is used to monitor the development of the fetus during pregnancy. The assay used in determining urine estrogen is the kober reaction.
This method involves the heating of a urine sample in a strong aqueous solution containing hydroquinone. After cooling and dilution the reddish brown solution is then measured for its absorbance.
Other color reactions ...
click here
Note:
Use of ampicillin or neomycin causes a decrease of urine estrogen because of the diminished hydrolysis of estriol conjugated. This is caused by the antibiotic that reduces the bacterial level in the urine.
Meprobamate, L-dopa, and phenolphthalein gives falsely elevated levels of estrogen in urine.
Immunoassay techniques
ELISA estradiol test
Measurement of estradiol in serum makes use of ELISA just like the progesterone tests. It employs the principle of competitive binding where the patient's estradiol competes with the standard control.
Then extra unbound estradiol is washed and then TMB is added to make a color blue solution. It is then incubated and then stop solution is added and then measured spectrophotometrically in 450 nm.
Precautions
Oral contraceptive and some vitamins interfere with progesterone and estrogen testEstrogen and progesterone levels vary from different menstrual phase
Glucose level should be tested before performing the test
Universal precaution must be observed
Avoid hemolysis of specimen
Normal values:
Progesterone
1st 3 months :9-50 ng/ml
2nd 3 months : 18-150 ng/ml
3rd 3 months : 60-260 ng/ml
male: 0.1-0.3 ng/ml
luteal phase: >10 ng/ml
Estrogen
Adult women: 30-400 pg/ml
Menopaused women: 0-30 pg/ml
Males: 10-50 pg/ml
click here for serum progesterone and estrogen reference range.
Photo Credits
http://www.lifesure.co.in/precaution.htm
http://www.herbalinfusions.ca/blog/category/tea-health/page/2/
http://trgmeetings.com/site/index.php?option=com_content&view=article&id=3&Itemid=3
http://www.flickr.com/photos/dubai_poeple/282667447/
