BY,

SANA ROBAB
Widal test is a serological test that is done for the diagnosis of
typhoid fever caused by Salmonella organism.
This test detects the “O” and “H” antigen of Salmonella typhi and
paratyphi A,B and C.
When facilities for culturing are not available, the Widal test is the
reliable and can be of value in the diagnosis of typhoid fevers in
endemic areas.
This was developed by Georges Ferdinand Widal in 1896
Typhoid fever or enteric fever occurs when S. Typhi, S. Paratyphi
A, S. Paratyphi B, S. Paratyphi C infect the human body.
Body responds to this antigenic stuimulus by producing
antibodies whose titre rise slowly in early stages, to a maximum
and then slowly falls till it is undetectable.
 Persons with typhoid fever carry the bacteria in their
bloodstream and intestinal tract.
Transmitted through the ingestion of food or drink contaminated
by the feces or urine of infected people.
How Widal Test
Is Done:

can be done in two

ways,
1)slide method
2)tube method
The patient’s serum is tested for O and H antibodies (agglutinins)
against the following antigen suspensions (stained suspensions):
1)S. Typhi 0 antigen
2)S. Typhi H antigen
3) S. Paratyphi A H antigen
4) S. Paratyphi B H antigen
5) S. Paratyphi C H antigen
Note: “H” suspensions are colored red while “O” suspensions color
is “Blue”.
REQUIREMENTS:
1)Reagents commercially prepared
antigen suspension of
I. S. typhi O (2-8°C)
II. S. typhi H (2-8°C)
III. S. paratyphi AH (2-8°C)
IV. S. paratyphi BH (2-8°C)
V. Positive control (2-8°C)
VI. Negative control (2-8°C)
2) Glass slide/test card
3)Disposable mixing sticks
1)Place one drop of positive control on one reaction circles of the
slide.
2)Pipette one drop of Isotonic saline on the next reaction cirlcle. (-ve
Control).
3)Pipette one drop of the patient serum tobe tested onto the
remaining four reaction circles.
4)Add one drop of Widal TEST antigen suspension ‘H’ to the first
two reaction circles. (PC & NC).
5)Add one drop each of ‘O’, ‘H’, ‘AH’ and ‘BH’ antigens to the
remaining four reaction circles.
6)Mix contents of each circle uniformly over the entire circle with
separate mixing sticks.
7)Rock the slide, gently back and forth and observe for agglutination
macroscopically within one minute.
When the colored, smooth, killed Salmonella antigen suspensions
are mixed/incubated with patient serum, anti salmonella antibody
present in the patient serum react with the antigen suspensions to
give agglutination.
Agglutination is a positive test result, indicating presence of anti
salmonella antibodies in the patient serum.
No agglutination is a negative test result indicating absence of anti
salmonella antibodies.