PREGNANCY TESTS
• From the earliest stage development(say 9 days) the placenta produces
hormones either on its own or in conjunction with the fetus.
• The placenta trophobast produces appreciable amounts of
hormone;Human chorionic gonadotropin(HCG) that is excreted in
urine.
• HCG is not found in the urine of normal ,young , non-pregnant
women.
• Increased urinary levels of HCG form the basis of most tests of
pregnancy and for trophoblastic tumor in men.
• All pregnancy test are designed to detect HCG.HCG is present in
blood and urine wherever there is living chorionic/placental tissue.
• HCG can be detected in the urine of pregnant women 26 to 36 days after the first
day of last menstrual period or 8 to 10 days after conception.
• Pregnancy tests should be negative 3 to 4 days after delivery.
• There are several methods of testing pregnancy even at very early stages either
using female rats or Zenopose frogs , all of these have been supplemented by the
immunological tests that are easier to perform.
• All of them depend on the presence of HCG in the urine of pregnant mothers.
• In certain disease condition a similar hormone may be produced in males.
• Tests can be grouped into
1. Biological
2. Immunological
3. Radiological(Ultrasound/ultrasonography)
1. BIOLOGICAL DIAGNOSTIC TESTS
Involves injection of urine into various animals.
It is time consuming and costly and 99% accurate
Examples of the tests:
a. Aschheim-Zondek mouse test
• Urine is injected subcutaneously into immature mice
• Appearance of blood filled ovulated follicles on 5 day
confirms pregnancy
b. Friedman rabbit test
• Intravenous injection of urine into virgin female rabbits
causes ovulation in 18 hrs.
c.Galli mainini frog test
• Injection of urine into the dorsal lymph sac of male frogs or toads,
which cause shedding of sperms in about 3hrs
d. Hogben test
• Adult female toads are used in this test
• Injection of urine into the lymph space causes ovulation within 18 hrs .
2. IMMUNOLOGICAL TESTS
HCG secreted by syncytiotrophoblast cells of the
placenta is antigenic
Antibodies can be produced against the hormones by
injecting into rabbits
Antibodies are available commercially and are
employed to detect presence of HCG by;-
Precipitation.
One step immunoassay test.
PRECIPITATION TEST
• Precipitation Reaction is a type of
antigen-antibody reaction, in which the antigen occurs
in a soluble form. When a soluble antigen reacts with
its specific antibody, at an optimum temperature and
PH in the presence of electrolyte antigen-antibody
complex forms insoluble precipitate.
One step immunoassay test
Principle : Use antibody that will specifically bind to the antigen of
interest based. The test is based on the combination of monoclonal
antibody dye conjugated with polyclonal solid phase antibodies for
qualitative detection. Monoclonal antibody dye conjugated is specific
on to HCG hormones while Polyclonal solid phase antibodies is non
specific.
• In presence of HCG a pink purple colored band develops
• A control is provided to check the potency of the test reagent
HOW TO TAKE URINE SAMPLE
Restrict taking water for 12-14hrs
Urine collected in clean container in the morning
It should be free from protein and blood
PROCEDURES
1. Remove the test strip from the sealed pouch
2. With arrows pointing toward the urine specimen , immerse the
strip vertically in the urine at least for 10 seconds , do not pass
the maximum line (MAX) on the test strip when immersing the
strip
3. Place the strip on the non-absorbent flat surface , start the timer.
4.Report should be read in 5 minutes
5.Interpretation
CAUSES OF FALSE POSITIVE RESULTS
Excessive protein /blood in urine
At menopause
At the time of ovulation due to increased secretion of Lutenizing hormone(LH)
Benign /malignant of the placenta
Drugs such as thiazide diuretics and steroids may affect early pregnancy test
CAUSES OF FALSE NEGATIVE RESULTS
HCG level too low
Testing too early/soon
Ectopic pregnancy
Advantages of Immunological test
Less costly
Simpler
Easy to carry out
Take very little time in reporting
Can confirm pregnancy within ten days of conception
D. Radio-immunoassay
• HCG radiolabeled with iodine treated with fixed amounts of antibodies and mixed
with urine
• More sensitive and can detect low concentrations of HCG
E. ELISA (Enzyme-linked immune sorbent assay
• Same as RIA except
• An enzyme is used in place of radioactive substance
• Enzyme acts on the substrate to produce blue colour which is a positive test for
pregnancy.
3. ULTRASONOGRAPHY
Most reliable
Gestational ring is evident as early as 5 th week of
pregnancy ,cardiac pulsation by 10 th week, and fetal
movements by 11th week.
