Chapter
Pregnancy Tests
86
INTRODUCTION
BIOLOGICAL TEST
IMMUNOLOGICAL TESTS
INTRODUCTION urine is injected subcutaneously into immature rat and
ovarian changes are observed after 6 hours. If the
Pregnancy test is the test used to detect or confirm urine is injected intraperitoneally, the ovarian changes
pregnancy. The basis of pregnancy tests is to determine can be observed within 2 hours.
the presence of the human chorionic gonadotropin
(hCG) in the urine of woman suspected for pregnancy. FRIEDMAN TEST
Both biological and immunological tests are available
In this test, 10 to 15 mL of urine is injected intravenously
to determine the presence of hCG in the urine of the
into rabbit and ovulation is observed by examining the
pregnant woman. ovaries after 48 hours.
BIOLOGICAL TESTS HOGBEN TEST
These tests are performed by using experimental animals. In this test, about 20 to 30 ml of urine is concentrated
The biological tests for pregnancy can be performed only and injected into the dorsal lymph sac of South African
after 2 or 3 weeks of conception so that, the concentration toad, Xenopus levis. If hCG is present in the urine, it
of hCG in urine is sufficient to show the result. causes ovulation after 12 hours.
ASCHHEIM-ZONDEK TEST GALLI-MAININI TEST
Aschheim-Zondek test was the first test invented for In this test, 2 mL of urine is injected into the male
confirming the pregnancy. It depends upon the ovarian amphibian (toad or frog). hCG in urine causes expulsion
changes in immature mice caused by hCG. The of spermatozoa within 2 hours.
immature mice do not ovulate naturally. Biological tests are outdated after the development
of immunological tests.
Ovulation occurs only if hCG is injected. 2 mL of urine
from the woman suspected for pregnancy is injected daily
Disadvantages of Biological Tests
for 2 days into the immature mice. 5 days after injection
of urine, the mice are killed. The ovaries are examined for Biological tests for pregnancy are replaced by
the presence of corpora lutea (plural for corpus luteum) immunological tests because of several disadvantages:
and hemorrhages, which indicates ovulation. Ovulation 1. The biological test require animals
is due to the presence of hCG in urine. 2. Tests can be performed only after 2 to 3 weeks
of pregnancy so that sufficient quantity of hCG is
KUPPERMAN TEST excreted in urine
3. Results are not obtained quickly; one has to wait for
Kupperaman test is the modification of Aschheim- 2 to 48 hours
Zondek test, in order to save time. In this, an immature 4. Tests involve tedious procedures such as sacrificing
rat is used instead of immature mice. About 2 mL of the animals.
Chapter 86 t Pregnancy Tests 509
IMMUNOLOGICAL TESTS
Presence of hCG is also determined by using
immunological techniques. Immunological tests
are based on double antigen-antibody reactions.
Commonly performed immunological test is known as
Gravindex test.
PRINCIPLE
Principle is to determine the agglutination of sheep
RBCs coated with hCG. Latex particles could also be
used instead of sheep RBCs.
REQUISITES
FIGURE 86.1: Immunological test for pregnancy
1. Antiserum from Rabbit
Urine from a pregnant woman is collected and hCG is
isolated. This hCG is injected into a rabbit. free antibody is available. Agglutination of hCG molecules
The rabbit develops antibodies against hCG. The by antibodies is not visible because it is colorless.
antibodies are called hCG antibody or anti-hCG. The Later when latex particles are added, these
rabbit’s blood is obtained and serum is separated. particles are not agglutinated because, free antibody
The serum containing hCG antibody is called rabbit is not available. Thus, absence of agglutination of latex
antiserum or hCG antiserum. It is readily available in particles indicates that the woman is pregnant.
the market.
Presence of Agglutination of Latex Particles
2. Red Blood Cells from Sheep If urine without hCG is mixed with antiserum, the
RBCs are obtained from sheep blood and are coated with antibodies are freely available. When latex particles
pure hCG obtained from urine of the pregnant women. are added, the antibodies cause agglutination of these
Nowadays, instead of sheep RBCs, the rubberized latex particles. Agglutination of latex particles can be
synthetic particles called the latex particles are used. seen clearly even with naked eye. Thus, presence of
agglutination of latex particles indicates that, the woman
3. Urine is not pregnant (Fig. 86.1).
Fresh urine sample of the woman, who needs to confirm
pregnancy is used for Gravindex test. ADVANTAGES OF IMMUNOLOGICAL
TESTS FOR PREGNANCY
PROCEDURE 1. Immunological tests are more accurate
1. One drop of hCG antiserum is taken on a glass 2. Result is obtained quickly within few minutes
slide. One drop of urine from the woman who wants 3. These tests can be carried out very easily. The
to confirm pregnancy is added to this and both are procedure is not cumbersome, as in the case of
mixed well. biological tests
2. Now, one drop of latex particles is added to this and 4. Immunological tests can be performed on 5th day
mixed. of conception. By biological methods, the tests can
be performed only after 2 or 3 weeks of conception.
OBSERVATION AND RESULT It is because, the concentration of hCG required
for producing changes in the animals is excreted in
Result is determined by observing the agglutination of urine only after 2 or 3 weeks of pregnancy
latex particles added to mixer of hCG antiserum and 5. Recently available immunological tests are more
woman’s urine. sensitive and involve single step method. Test kit
is available in the form of cards. These pregnancy
Absence of Agglutination of Latex Particles test cards can be used even in the first few days
If hCG is present in urine, it is agglutinated by antibodies of conception. Most sensitive test can detect hCG
of antiserum and all the antibodies are fully used up. No level as low as 20 mIU/mL.
