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Pregnancy Test Detecting Human chorionic gonadotropin in urine
BCH 472
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Human chorionic gonadotropin
• Human chorionic gonadotropin (hCG) is a glycoprotein hormone produced by a
portion of the placenta following implantation.
• The qualitative hCG test can be used to see if a woman is pregnant or not.
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hCG Role in pregnancy:
• Promotes the maintenance of the corpus luteum during the beginning of
pregnancy in the ovary . This allows the corpus luteum to secrete the
progesterone during the first trimester. Progesterone enriches the uterus with
a thick lining of blood vessels and capillaries so that it can sustain the
growing fetus.
• Human chorionic gonadotropin also plays a role in cellular
differentiation/proliferation.
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hCG level
• During the first trimester, hCG levels rise steadily and rapidly, peaking
around 10 weeks’ gestation, and subsequently taper off to less than 10% of
peak levels.
• By the end of the third month it has reached a low level which will remain
constant for the duration of the pregnancy.
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Notes:
• In pregnant women, depressed urine hCG levels may indicate threatened
abortion .
• Some cancerous tumors produce this hormone; therefore, elevated levels
measured when the patient is not pregnant can lead to a cancer diagnosis.
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Practical Part
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Objective:
To detect and confirm pregnancy.
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Principle:
• Urine pregnancy tests use the enzyme-linked immunosorbent assay (ELISA)
technique, using a highly specific monoclonal antibody directed against the -
subunit of human chorionic gonadotropin (-hCG).
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Principle: Pregnancy test strip consist of :
• The reaction zone: contains soluble anti-hCG antibody-Enzyme. These are mouse monoclonal antibodies linked to an enzyme.
• The test zone: contains immobilized polyclonal mixture of hCG- antibody + dye substrate.
• The control zone: the dye substrates + anti-(anti-HCG Ab-Enzyme conjugates) Ab can recognize epitopes on the mouse monoclonal .
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• http://www.sumanasinc.com/webcontent/animations/cont
ent/pregtest.html
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Sequence of events:
• A few drops of urine is transferred to the specimen well (loading well).
• Urine will flow by capillary action from loading well towards zone R carrying along with it the HCG
hormone.
• At R zone, the HCG hormone will react and bind with the soluble anti-HCG Ab-enzyme conjugates
forming a complex of HCG hormone - HCG Ab - enzyme conjugate.
• This complex will migrate towards zone T.
• At zone T, this complex will react and bind with the immobilized anti-HCG Ab, once it binds with the
immobilized Ab, this will activate the enzyme thus allowing to act on the dye substrate and produce a color
that indicates a positive pregnancy result.
• The excess free HCG Ab - enzyme conjugates will pass from zone T to zone C.
• At zone C, this free HCG Ab - enzyme conjugates, will react and bind with the immobilized anti-(anti-HCG
Ab-Enzyme conjugates) Ab there, once bound it will activate the enzyme, thus allowing to act on the dye
substrate and produce the color detecting at zone C which is an indicator of the activity or reliability of
the test.
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At R zone
At T zone
At C zone
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Pregnant (+) Non pregnant (-)
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Specimen Collection and Preparation
• Collect at least 1 mL of urine in a clean, dry, plastic or glass container with
no preservatives.
• Specimens may be collected at any time of the day, however the first
morning sample generally has the highest concentration of HCG and is the
specimen of choice.
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Procedure: • Holding a Sample Dropper vertically, add exactly four drops of the urine
specimen to the sample well.
• Read results at time indicated in procedure.
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Results
• Follow the instructions on the reagent package insert provided by the
instructor to properly perform the test.
Interpretation of Results:
• Based on the package insert correctly interpret the results of the pregnancy
test on the patient samples
• Record results as “Positive” or “Negative”
RESULT SAMPLE TESTED
Urine sample
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Pregnancy test strip
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Discussion
Comment on the results and state if sample is pregnant .