WIDAL

AGGLUTINATION
TEST
dr. Dewi Suryani, M.InfectDis (MedMicro

CONTENT
Principle

of Agglutination Test
Widal Agglutination Test
Performance technique
Interpretation of test results
- false positive
- false negative

Basic Principle of
Agglutination Test
Agglutination is a
classic serologic
reaction that results
in clumping of a cell
suspension by a
specific antibody,
directed against
specific antigen

Image: Mayer, 2010

Image: Serum Analysis

Centre, n.d
Source: Olopoenia & King,
2000

History of Widal
Agglutination Test
Widal Agglutination
test was first
developed by
Georges Fernand
Isidore Widal
(French physician
and bacteriologist)
in 1896 to utilse the
diagnose
of Typhoid
Source: Rao, 2009
Fever.

Image: APPL, 2008

Different type of Detection

Methods
1. Serology Ab
detection:
Widal

2.
Immunochromatogrph
y essay: Ag
detection w/
monoclonal and
polyclonalAb

Different type of Detection

Methods
3. PCR to detect S. Typhi

3. Culture

Widal Agglutination Test

The Widal Test was based on
demonstrating the presence of agglutinin
(antibody) in the serum of an infected
patients, against the Antigens of
Samlomenella typhi and Salmonella
paratyphi A and B:
H (flagellar) Antigens
O(Somatic) Antigens
Vi (Virulen) Antigens

SLIDE TEST

Performance technique:
qulitative
Make 6 wells/circle on object

Performance techniques:
Semiquantitative

Circ
le
no

Serum
volume
(ml)

Antigen
drop

Titre

0.02

1 drop

1:80

0.01

1 drop

1:160

0.005

1 drop

1:320

0.0025

1 drop

1:640

Performance technique:
Semi Quantitative

Performance technique

videoplayback_3.FLV

INTERPRETATION
Serologic

studies are considered

significant in endemic area if
patients have four-fold or greater
increases in O or H agglutinin titres
in serum specimens obtained several
days apart

Source: Olopoenia & King,

2000

INTERPRETATION
Report

of test should be:

Agglutination Agglutination +, with titres
This is significant as there could be
misleading interpretation due to false
positive and false negative results.
Source: Olopoenia & King,
2000

REMARKS
A

rising titre is more significant then

a single high titre
There are range of cut off point for
titres --> may vary between
population and need to be
established for each area
Always correlate clinical findings with
lab findings

Interpretation of Widal
Test
False Positive

False Negative

Previous immunisation with

Salmonella antigen

An inadequeate inoculum of
bacetrial antigen in the host to
induce antibody production

Cross-reaction with non

typhoidal Salmonella

technical difficulty or errors in

the performance of the test

variability and poorly

standarised commercial antigen
preparation

previous antibiotic treatment

Infection with malaria or other

Enterobactericaea

variability in the preparation of

commercial antigens

other disease such as dengue

read more then 1 minute after
mixing the slide test
Source: Olopoenia & King,
2000

References
Mayer G. Immunoglobulins-Antingen-Antibody Reactions and
Selected Test. 2010 [updated 12 july 2010; cited 1 Novemeber ].
Available from: http://pathmicro.med.sc.edu/mayer/ab-ag-rx.htm.
Olopoenia LA, King AL. Widal Agglutination Test - 100 Years Later:
Still Plagued Controversy. Postgrad Med J. 2000; 76.
Rao S. Widal Test. Davanere; 2009 [updated 2 November 2011].
Available from: www.microrao.com.
Serum Analysis Centre. Serology. Kolkata; n.d [updated 2
November ]. Available from:
http://www.indiamart.com/serum-analysiscentre-privatelimited/se
rology.html
.