Hormonal Assays
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Transcript of Hormonal Assays
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Hormonal Assays
Dr. Saleh AhmedAss. Prof. of Tumor Biology &
BiochemistryFaculty of Medicine
Umm Al Qura Univ. & Ain Shams Univ.
By
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Some selected hormonal assays
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Topics
PRL TSH LH FSH E2
ELISA Technique
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PRL Prolactin Lactogenic hormone Lactogen
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PRL assay measure the concentration of the prolactin hormone in the blood.
PRL is a polypeptide hormone secreted by lactotrophs of the anterior pituitary gland, primarily for the development of mammary glands for lactation during pregnancy and for stimulating and maintaining lactation.
Reference values:• Premenopuasal: < 20 ng/ml• Postmenopausal: < 12 ng/ml
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Conditions for PRL assay Patient has to limit physical activity 12
hrs before test. Fasting for 12 hrs before test. Patient has to avoid stress, or
stimulation for 30 minutes before test. Sample drawn in the morning (3 - 4 hrs)
after awakening. Handle sample gently to prevent
hemolysis.
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PRL is under the complex regulatory system of estrogen, progesterone, dopamine, and thyrotropin-releasing hormone (TRH).
The function of PRL in males is unknown.
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PRL levels are measured in the workup of galactorrhea, amenorrhea, infertility, impotence, and in cases of suspected pituitary tumor.
An elevated PRL classically presents with the syndrome of galactorrhea-amenorrhea in women, and the syndrome of infertility-impotence in men.
Men with elevated PRL typically have a low serum testosterone. However, testosterone replacement alone will not reverse the symptoms, the PRL must also be reduced.
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Hypersecretion Hyposecretion
Physiologic
Pharmacologic
Pathologic
Rare, if happened this may be due to pituitary necrosis or infarction
Clinical significance of PRL level
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Pathologic Pharmacologic PhysiologicHypothalamic disease Methyldopa Pregnancy
PRL secreting tumor Reserpine Lactation
Hpothyroidism Cimetidine Excerise
Addsion’s disease Estrogen Eating
Chronic renal failure Morphine Stress
Cirrhosis
Causes of Hyperprolactinaemia
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PRL & Pituitary tumor
?? 60% pituitary tumor > 100 ng/ml
?? 90% pituitary tumor > 200 ng/ml
100% pituitary tumor > 300 ng/ml
Modest elevation can be associated with pituitary tumor
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Hyperprolactinemia may clinically present as:
Amenorrhea Galactorrhea Infertility Osteoprosis Impotence Erectil dysfunction Infertility
Females
Males
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PRL High (> 23 ng/ml) Normal (< 23 ng/ml)
TSH
Normal High
MRI or CT Hypothyroidism
Normal hyperplasia Microadenoma Macroadenoma
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TSH Thyroid Stimulating Hormone Thyrotropin
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Secreted by the thyrotrophic cells
of the anterior pituitary.
It stimulates the growth of the
thyroid follicular cells & step by step
thyroid hormone synthesis
TSH
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TSH valueThe best single test for the thyroid function
I Screening for thyroid dysfunction
a- TSH decreased with hyperthyroidism
b- TSH increased with hypothyroidism
II Monitoring thyroid replacement therapy
(eg. Levothyroxine)
III Monitoring anti-thyroid therapy
(eg.Propylthiouracil, methimazole or
radioactive iodine)
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The ultra-sensitive assay in contrast to the older TSH assay which was unable to distinguish patient values in the normal range from those which were abnormally low.
It is the feeling of thyroid specialists that measurement of the TSH, complemented by FT4 measurement, represents the best and most efficient combination of blood tests for the diagnosis and follow-up of most patients with thyroid disorders.
TSH value
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There has been a revolution in the approach to thyroid testing as the result of the development of ultra-sensitive TSH assays.
1st generation tests were able to measure levels of TSH to 1 IU/mL.
2nd generation tests were able to measure levels of TSH to 0.1IU/mL
3rd generation tests can measure TSH to 0.01IU/mL, a point at which hyperthyroidism may be diagnosed in ill patients.
4th generation assay able to measure TSH to 0.001IU/mL
TSH value
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Follow up patients on thyroxine supplementation (eg. Thyroxine or Synthroid) the TSH is an appropriate single test that can be followed and used to determine need for adjusting dosing.
TSH is an indication to increase the thyroxine dose.
TSH indicates a need to decrease the thyroxine dose.
Normal TSH range is the goal for patients on supplementation.
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Clinical conditions associated with thyroid dysfunction
Amenorrhea
Oligomenorhea
Anovulation.
Inadequate corpus luteum.
Subfertility
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Ultra-sensitive TSH
High Normal Low
FT4 Normal Thyroid FT4
Low Normal High Normal
Hypothyroidism
SubclinicalHypothyroidism
Hyperthyroidism
FT3
Normal High
SubclinicalHyperthyroidism
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LH & FSH
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They are secreted by the anterior pituitary.
The alpha subunit is identical for all glycoprotein hormones (TSH, HCG, LH & FSH), but the beta subunit differs.
The peak of FSH is coincident with the peak of LH, but it is of lesser magnitude & briefer duration.
Following the mid-cycle surge of LH & FSH, there is drop in both.
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• In female patients, indicate the phase of the menstrual cycle or duration of menopause on the lab request.
• Medications containing estrogen and progesterone should be discontinued 4 weeks before test.
Conditions for the detection of LH & FSH
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The clinical utility of testing LH & FSH levels, includes:• Evaluation of menstrual disorders.• Aids in the diagnosis and treatment of
infertility– To evaluate ovarian reserve of egg supply
in females– To evaluate low sperm count in males
• Assists in the detection of ovulation and monitors therapy to induce ovulation
• Evaluation of failure of sexual maturation in adolescence.
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The clinical utility of testing LH levels, includes:
• Assists in distinguishing between primary (ovarian or testicular) and secondary (pituitary or hypothalamic) gonadal failure or hypogonadism.
• Evaluation of impotence in males.
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E2
Estradiol-17 beta Estrogen fraction Serum Estradiol
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Estrogens
More than 30 estrogens have been identified, only 3 estrogens are used in clinical practice: estrone (E1), estradiol (E2), estriol (E3).
E2 is the primary form of estrogen in women, and is responsible for development of secondary sexual characteristics.
E2 is produced by the ovaries in small basal amounts prior to the onset of puberty
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Gonadotropin releasing hormone (GnRH) stimulates the pituitary to produce LH and FSH.
FSH stimulates ovarian follicle formation and estrogen secretion; LH triggers follicular rupture and ovulation.
E2 secretion cause changes in the cervical mucus secretion to optimize fertility.
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Potential use of E2 measurement
Delayed Puberty
Precocious Puberty
Secondary Amenorrhea
Breast Cancer Risk
Infertility
Gynecomastia in males
Females
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E2 rangeConv. Units (pg/mL) Phase
20-150 Females (Adults) Early follicular100-400 Females (Adults) Late follicular150-750 Females (Adults) Midcycle peak50-250 Females (Adults) Luteal
Up to 35,000 Females (Pregnant)<50 Females (On OCPs)<30 Females (Post-menopause)
30-280 Females (Pubertal)<15 Females (Pre-puberty)
10-60 Males (Adults)<10 Males (Prepubertal)
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ELISA
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ELISA Enzyme-Linked Immunosorbant Assay
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Enzyme-Linked ImmunoSorbent Assay Biochemical reaction to identify the
presence of antigen or antibodies in a sample Direct: Antigen detection Indirect: Antibodies detection
ELISA
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Non-radioactive High specificity Color change or florescence allows to use
photometric measurement. Amplification of minute levels allows for
high sensitivity Qualitative and Quantitative
Advantages of ELISA
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Direct ELISA: Antigen detection
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Indirect ELISA: Antibodies detection
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Micro-particle Enzyme Immuno Assay
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MEIA
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MEIA
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ELISA assaysThyroid Stimulating Hormone (TSH)
Hormones
Thyroxine (T4) & Triiodothyronine (T3)Free Thyroxine (fT4) &Free Triiodothyronine (fT3)CortisolFollicle-Stimulating Hormone (FSH)Luteinizing Hormone (LH)ProlactinEstrogenProgesteroneTestosteroneHuman chorionic gonadotropin (hCG)
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ELISA assays
Troponin ICardiac Markers
Myoglobin
Creatinine Kinase-MB (CK-MB)
B-type Natriuretic Peptide (BNP)
High Sensitivity CRP (hsCRP)
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ELISA assays
Hepatitis A virus (HAV) and its antibodiesHepatitis Markers
Hepatitis B virus (HBV) and its antibodies
Hepatitis C virus (HCV) and its antibodies
Hepatitis D virus (HDV) and its antibodies
Hepatitis E virus (HEV) and its antibodies
HIV-1 HIVHIV-2
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ELISA assays
(hCG) Hormones
Tumor markers
Alpha Fetoprotein (AFP)Carcinoembryonic Antigen (CEA) Prostate Specific Antigen (PSA)
Oncofetal Antigens
CA 15-3CA 549CA 27-29CA 125
Carbohydrate Markers
CA 19-9CA 50CA 72-4CA 242
Blood Group Antigens
Monoclonal IgA, Monoclonal IgGMonoclonal IgM, B2-Microglobulin Proteins
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ELISA assays
Rub IgG and Rub IgMCongenit
al Factors
Toxo IgG, Toxo IgM
CMV IgG, CMV IgM
Ferritin
Metabolic TestsVitamin B12
Folate
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ELISA assays
Anti-CCP(anti-cyclic citrullinated peptide antibody)
Specific Proteins
Apoliprotein A1 (Apo A1)
Apoliprotein B (Apo B)
Complement C3 & C4 (C3, C4)
Immunoglobulin G (IgG)
Prealbumin
Rheumatoid Factor (RF)
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