Widal Test

Laboratory diagnosis
The diagnosis of S. typhi infection is based on
primarily on detection of S. typhi (antigen) in culture
and remains the standard technique for diagnosis of
typhoid fever
The widal agglutination test & the
counterimmnonuelectrophoresis assay are commonly
used in developing countries to aid in the diagnosis of
S. typhi
Widal Agglutination test
It also referred as febrile aggn test.
It was introducef as serological test by F. Widal (1896)
to aid in the diagnosis of typhoid fever
This test uses suspension of killed S. typhi as Ag to
detect anti-S. typhi Ab in px serum sample by using an
aggn as an endpoint
Ag suspension for the Widal aggn test
The widal aggn test was developed to measure the
prsence of anti O & anti H Ab in the serum of px
infected w/ S. typhi, using bacterial suspensions of
killed S. typhi & S. paratyphi B that have been
treated so that they retain only the O & H Ag.
S. paratyphi A & S. pratyphi B share O Ag w/ S. typhi
& both of these organisms cause diarrhea & enteric
fever.
Ag suspension for the Widal aggn test
During the course of the infection, IgM anti O Ab are
detected early in the infection. While the anti H IgG
Ab develops later but persist for a longer period of
time.
Approaches to the widal test
This test can be performed in test tubes or on slides,
using commercially available Ag suspension.
The tube technique is a more labor-intensive approach
than the slide test in that it requires ten serial dilutions
for each Ag.
Therefore, it is only used to confirm the results of a
slide test
Slide technique
Introduced by Welch & Mickle in 1936
It can performed on a large plate of glass that is
divided into squares.
Different volumes (e.g. 80, 40, 20, 10, 5) of clear,
undiluted & unheated serum are added to specific
squares on the glass or card.
And add a constant volume & conc. of Ag suspension
to each square.
 After adding a drop of the prepared Ag to each clinical
sample, the contents of each square are mixed using a
glass rod (or wooden applicator)
Beginning w/ the sample that is most diluted.
The glass plate is rocked & tilted for a period not to
exceed 3 minutes
Depending on the extent of aggn, a strong light source
may be required for visualization
Aggn is visualized as clumps
Reporting results
The results are scored from 0 to 4+
Where 0 is represents no aggn
When the aggn is present, the results are graded +1,
+2, +3, +4 w/c represents 25, 50, 75, & 100 % aggn
respectively.
A single rapid slide test is not considered positive
unless a value of at least +2 is assigned to a sample
taken in the first 2-3 weeks of infection
Reporting results
False negative readings arise if the test is performed
too soon during the course of infection, because Ab
will not have developed to sufficient titers to be
detected.