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
.