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Widal Test

The Widal test is a tube agglutination test used for diagnosing enteric fever by detecting antibodies against Salmonella typhi and S. paratyphi in patient serum. It requires specific antigen suspensions and involves mixing serum dilutions with antigens, followed by incubation and reading for agglutination. Significant results include an O titre of 1:100 or more and an H titre of 1:200 or more, with a four-fold rise in titres over 7-10 days being more diagnostic.

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5 views1 page

Widal Test

The Widal test is a tube agglutination test used for diagnosing enteric fever by detecting antibodies against Salmonella typhi and S. paratyphi in patient serum. It requires specific antigen suspensions and involves mixing serum dilutions with antigens, followed by incubation and reading for agglutination. Significant results include an O titre of 1:100 or more and an H titre of 1:200 or more, with a four-fold rise in titres over 7-10 days being more diagnostic.

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AIM: To study Widal test.

PRINCIPLE: It is a tube agglutination test employed for the serological diagnosis of enteric fever, in which
antibodies (agglutinins) against Salmonella typhi, S. paratyphi A and B infections are detected in patient
serum. Salmonella antibodies start appearing in the serum at the end of first week and rise sharply
during the third week of enteric fever. Two specimens of sera at an interval of 7 to 10 days are preferred
to demonstrate a rising antibody titre.

A total of 4 antigen suspensions namely TO, TH, AH and BH are employed in the test.

REQUIREMENTS: Widal rack, Dreyer's tubes for 'H' agglutinins (conical bottom), Felix tubes for 'O'
agglutinins (round bottom), Patient serum, antigen suspensions, physiological saline, pipettes, waterbath
at 37 °C.

PROCEDURE:
Equal volume (0.4 ml) of serial dilutions of the serum (1:10 to 1:640) and the H and O antigens are mixed
and incubated in a water bath at 37 °C for 4 hours and read after overnight refrigeration at 4 °C. Control
tubes containing the antigen and normal saline are included to check for autoagglutination. H
agglutination leads to the formation of loose, cottonwool clumps, while O agglutination appears as a
granular deposit at the bottom of the tube. The highest dilution of serum showing agglutination (carpet
formation) indicates the antibody titre against that particular antigen. Control tubes show a compact
deposit (button formation).

INTERPRETATION:

O titre of 1:100 or more and H titre of 1:200 or more is significant.

The demonstration of a four-fold or greater rise in titre of both H and O antibodies at an interval of 7-10
days is more meaningful and diagnostic than a single test.

In immunization with TAB vaccine, individuals may show high titres of antibody to S. typhi, S. paratyphi A
and B.

Test may also be positive in many healthy carriers.

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