SEROLOGY DEPARTMENT

Serology is the scientific study of serum and other bodily. In practice, the term usually refers to
the diagnostic identification of antibodies in the serum. Serological test may be performed for
diagnostic purposes when an infection is suspected, in rheumatic illness, and in many other
situations. Serology blood test help to diagnose patients with certain immune deficiencies
associated with the lack of antibodies.
Some tests classified under serology are: widal agglutination test, retroviral screening (RVS) test,
hepatitis B surface antigen (HBsAg) test, hepatitis c virus (HCV), pregnancy test, and syphilis test
(VDRL).
NOTE: All above mentioned tests under serology have the same procedures but different strips
kit, except for widal agglutination test.
PROCEDURE FOR SEROLOGY TEST (except for Widal)
 Spin the anti-coagulated blood sample in the bucket centrifuge at 12,000rpm for 5 minutes.
 Apply 2-3 drops of plasma/serum on the sample pad of the test strip with the pasteur pipette
(each test has its own strip).
 Read the result after 10 minutes.
RESULT
Double band (one on control and the other on test) = Positive
Single band (on the control alone) = Negative
No band = Invalid (to be repeated).

3.4.1 Widal Agglutination Test

The principle of Widal test is based on antigen-antibody reaction. The test is done to diagnose
Salmonella typhi which is the causative agent of typhoid.
Aim: To determine the presence of Salmonella typhi and Salmonella paratyphi in a patient’s
plasma.
Principle: The stained antigen suspensions are bacteria stained to enhance the reading of
agglutination test. The blue antigens are specific ‘O’ antigen while the red stained antigens are
specific to the flagella ‘H’ antigens.
Materials: Clean grease free tile, dry cotton wool, applicator stick, pasteur pipette, bucket
centrifuge, widal kits.
Procedure:
  A drop of plasma was applied on a clean grease free tile and a drop of each of the paratyphi
O, A-O, B-O, C-O, and H, A-H, B-H, C-H in square 1-8 was added into the plasma
respectively and was rocked gently for 3 minutes.
 Observe for agglutination.
Result: If the degree of agglutination shows 1:80 titre or above for both O and H anti-bodies, it
indicates positive result but if it has less than 1:80 titre, it indicates negative result.

3.4.2 Retroviral Screening (RVS) Test

Retroviral screening test is used is the diagnosis of the human immuno virus, HIV.
The virus can be transmitted through the following ways:
 Through transfusion of infected blood.
 Through the use of unsterilized surgical instrument.
 From mother to child during delivery.
 Through an unprotected sex with an infected person.

3.4.3 Hepatitis B Surface Antigen (HBsAg) Test

Hepatitis refers to an inflammatory condition of the liver. Hepatitis B is an infectious disease
caused by hepatitis B virus which affects the liver. It causes both acute and chronic infections. The
hepatitis B surface antigen (HBsAg) is most frequently used to screen for the presence of the
infection. It is the first detectable viral antigen to appear during infection. It is transmitted through
sexual intercourse, kissing and exchange of body fluids.
Principle: The human body produces a sufficient amount of antibodies against the hepatitis virus
within the plasma when the virus invades the body. The antibodies produced forms a complex
during reactions that can be seen as coloured bands on the test zone of the strip membrane
depending on the level of the antibodies produced which are proportional to the level of the
infection obtained. The antigen embedded strip reacts with the antibodies in the plasma. The result
is read in 10 minutes.

3.4.4 Hepatitis C Virus (HCV)

Hepatitis C virus (HCV) test is done to detect the presence or absence of Hepatitis C virus.
Hepatitis C is transmitted through direct contact with infected body fluids, typically through
injection drug use and sexual contact. HCV is among the most common blood-borne viral
infection. This virus affects the liver, causing inflammation which damages liver cells and
eventually fibrosis where the liver cells are replaced by tough, but non-functional fibrous tissue
and if left untreated can lead to cirrhosis of the liver. Symptoms of hepatitis C includes fatigue,
loss of appetite, fever, abdominal pain, jaundice (a yellowing of the skin and eyes), etc.
Principle: HCV test employs chromatographic lateral flow device in strip format. HCV antigens
are bond at the test (T) zone and anti-HCV monoclonal bodies are bond at the control (C) zone.
Upon plasma addition it migrates by capillary diffusion rehydrating the gold conjugate. If present
in sample, HCV Antibodies will bind with the gold conjugated antigens forming particles. These
particles will continue to migrate along the strip until the test zone where they are captured by the
HCV antigens generates a visible red line. The gold conjugate will continue to migrate alone until
it is captured in the control zone by the anti-HCV antibodies aggregating in a red line, which
indicates the validity of the test.

3.4.5 Veneral Disease Research Laboratory (VDRL) Test

Veneral Disease Research Laboratory (VDRL) test also known as syphilis test is a test done to
detect the presence or absence of Treponema palllidum bacteria in a patient’s plasma. Syphilis is
one of the most common sexually transmitted diseases (STDs). It is an infection caused by the
bacteria Treponema pallidum. Syphilis is transmitted through vaginal, oral, or anal sex with an
infected person. It is mostly through contact with a syphilis sore (chancre), a painless sore.
Symptoms include painless sore either in the anus, mouth or genitals, rashes, fatigue, weight loss,
fever and hair loss.
Principle: Recombinant syphilis antigen is immobilized in a test line region. After a plasma
sample is added, it reacts with syphilis antigen coated particles in the test. This mixture migrates
along the length of the test strip and interacts with the immobilized syphilis antigen.

3.4.6 Beta Human Chorionic Gonadotropin (β-hCG)

Beta-human chorionic gonadotropin (β-hCG) is a test that measures the amount of human
chorionic gonadotropin in the blood. This hormone is produced as soon as 10 days post-conception
and an above-normal level can confirm pregnancy.
Aside from this, the beta-hCG test may also be used during evaluations of fertility treatments (a
synthetic form of the hormone is sometimes used to help follicles mature and trigger ovulation),
as well as when there are concerns that something may be wrong with a pregnancy.
What Beta HCG Measures
Pregnancy testing involves the detection of hCG either in the urine or blood.
The urine test is a qualitative one in that it can only tell you if the sample is positive or negative
for hCG. The same goes for the qualitative hCG blood test.
In contrast, the beta hCG is a quantitative test, meaning it reveals not just that the hormone is
present in the blood, but in exactly what amounts. Levels of hCG are measured in milli-
international units per milliliter (mIU/ml).
The beta-hCG test is also used when there are concerns about pregnancy complications,
including miscarriage. In these situations, repeat tests may be performed every two to three days
to