TRUEchemie WIDAL(Antigen Agglutination) TEST KIT 2

8
8

(Stained Widal-Antigens for Slide Test) 2

for the qualitative and semi-quantitative determination of Widal in serum

INTENDED USE INTERPRETATION OF RESULTS:

The TRUEchemie Widal test for the qualitative and semi-quantitative determination of Widal The lowest volume of serum, which shows clear agglutination, indicates the cut off level of
Antibodies in serum. the positive test and the corresponding antibody titre as per the tube technique is given
below:
INTRODUCTION
Serum Volume Antibody Titre Serum Volume Antibody Titre
Salmonella typhi & Salmonella paratyphi are the causative agents of “Enteric Fever”. In
enteric fever, once the patient is on medication it becomes difficult to isolate the organisms. 0.08 ml 1: 20 0.01 ml 1:160
In serological tests the antibodies produced as a result of infection are detected by using
0.04 ml 1: 40 0.005 ml 1:320
the killed bacterial antigens. The antibodies from the patient’s serum react with the
corresponding antigens to cause clumping or agglutination. 0.02 ml 1: 80
The antigens of typhoid and paratyphoid consist of two distinct fractions-the stable somatic (C) TUBE TECHNIQUE USING SLIDE ANTIGENS:
‘O’ Antigen and the labile flagellar ‘ H’ antigen. The paratyphoid antigens are further a) Perform the assay for all four antigens or for that which has given a positive result in the
classified into A&B species. In typhoid and paratyphoid, the ‘H’ antigen is type specific
whereas the ‘O’ antigen is group specific. Screening Slide Test.
b) Take a set of six test tubes (10x75 mm) for each antigen. Dilute the serum sample and
Typhiscreen-S antigens are the standardised smooth suspension of killed bacterial
antigens for qualitative and semi-quantitative detection of [Link] and S. paratyphi set up the test as indicated in the table.
antibodies. The different color to each antigen facilitates the differentiation of antigens and
clear reading of agglutination. It also avoids the possible error of mixing and/or
misinterpretation. As undiluted serum is used in Slide Test, it is a simple, rapid and Tube No. 1 2 3 4 5 6
convenient screening test. The slide test antigens are standardised in such a way that they
can be used for either slide or tube technique. In doubtful cases, it is recommended to Dilution Saline 1:20 1:40 1:80 1:160 1:320
perform the tube technique for obtaining conclusive results. A marked rise in the titre to
one sero type (above 1:80) suggests infection. Diagnostically a rising antibody titre of at Control
least four fold (two tube difference) is considered more significant than a single test. It is
observed that individuals immunised with TAB vaccine may show a moderately high titre for Normal Saline 1.0ml 1.9 ml 1.0 ml 1.0 ml 1.0 ml 1.0 ml
all the antigens.

PRINCIPLE
Test Serum -- 0.1 ml
The antibodies present in the serum sample react with the corresponding bacterial antigens Diluted Serum -- -- 1.0 ml 1.0 ml 1.0 ml 1.0 ml
to give agglutination.
1.0 ml
PACK SIZE
discard

Kit Size 4 x 5 ml 2 + 2 x 5.0 ml

Cat. No. ADX841 ADX842
Kit Contents Appropriate one one one one one one
1) S. typhi “O” Antigen Reagent 1 x 5 ml 2 x 5 ml Antigen
2) S. typhi “H” Antigen Reagent 1 x 5 ml 2 x 5 ml drop drop drop drop drop drop
3) S. Paratyphi “AH” Antigen Reagent 1 x 5 ml -
4) S. Paratyphi “BH” Antigen Reagent 1 x 5 ml -
5) Widal Positive Control 1 x 0.5 ml 1 x 0.5 ml
Accessories 0

Widal Slide c) Mix well after each addition and incubate at 37 C for 16-20 hours.
d) Observe for agglutination. The highest dilution of Serum which shows clear - cut
STORAGE AND STABILITY
agglutination indicates the antibody titre.
0
All reagents are stable at 2-8 C till the expiry date mentioned on the labels. QUALITY CONTROLS
DO NOT FREEZE.
Positive & Negative Controls are used to validate the kit performance.
REAGENT PREPARATION INTERFERENCES
Ready to use reagents. In some non-infected cases, non-specific agglutinins may appear and react with the Febrile
antigens giving false results. Some vaccination may also produce agglutinins that react
SAMPLE / SPECIMEN AND STORAGE with Febrile antigens resulting in false results. Physicians should always evaluate all clinical
and laboratory findings before making a definitive diagnosis.
Fresh Serum.
0 NOTES
In case of a delay in testing, store at 2-8 C.
1. Sera from normal individuals may show agglutination up to 1:40 dilution.
2. Agglutination titre greater than 1:80 is considered significant and usually suggestive
WARNINGS AND PRECAUTIONS of infection.
3. Typhiscreen-S is only screening test. For confirmation of results, testing with
1. Bring all the reagents and samples to room temperature before use. Typhiscreen-T is recommended.
2. Shake all the antigens thoroughly before use. 4. The correlation of test results with typical clinical signs, symptoms and patien
3. Avoid using turbid, contaminated or inactivated serum. history should be taken into account before arriving at the final diagnosis.
5. As with all diagnostic procedures, the Physician should evaluate data obtained by
4. The disposal of the residues has to be done as per local legal regulations.
use of this kit in light of other clinical information.
TEST PROCEDURE
6. For accuracy of results, the procedure has to be followed meticulously.
(A) QUALITATIVE TEST (RAPID SCREENING SLIDE TEST):
REFERENCES
a) On a Slide with six circles, place 0.05 ml of test serum in each of the first four circles and
1. Cruickshank, R. (1982) Medical Microbiology, 12 th Edition, P.403.
0.05 ml each of Positive Control and Normal Saline in each of the last two circles 2. Felix, A. (1942) Brit. Med. J., 11, 597.
respectively.
b) Add one drop each of ‘O’, ‘H’, ‘AH’ and ‘BH’ antigens in the first four circles respectively
and one drop of any one antigen in the remaining two circles.
Consult instructions for use Catalogue number Use-by date
c) Mix the contents of each circle separately and spread it in the entire circle.
Consult instructions
For in vitro for use
diagnostic Catalogue number Use-by
Do not use if date
d) Rock the slide gently for One minute and observe for agglutination. Batch code package is
use only
damaged
INTERPRETATION OF RESULTS: For in vitro diagnostic Do not use if
8
Temperature limit 2-8 °C Batchfrom
Keep away codesunlight
2 use only Keep drypackage is
Agglutination with Positive Control and no agglutination with Normal Saline validate test damaged
results. No agglutination up to one minute is a negative test, and indicates the absence of 8
Temperature limit 2-8 °C
Manufacturer Date
Date of of manufacture
manufacture European
corresponding antibodies. 2 Conformity
EC REP Authorized
Keep dry
If device Warnings / Precautions
Agglutination within one minute is a positive test, and indicates presence of corresponding Keep away isfrom
non-sterile
sunlight ! Manufacturer Representative
NON
STERILE

antibodies. Then proceed for semi-quantitative slide or tube technique for determination of
antibody titre. Authorized
If device is non-sterile Warnings / EC REP
Representative
Do not observe result after One minute
NON
! Precautions
Effective date: 03.06.2019 Rev. B
STERILE

(B) SEMI QUANTITATIVE TEST: English version

a) Put one drop of normal saline in the first circle and 0.005 ml, 0.01 ml, 0.02 ml, 0.04 ml & Effective date: 02.05.2019 Rev. B
0.08 ml of test serum in the remaining five circles respectively. English Obelis
versions.a.
EC REP Bd General Wahis 53
b) To each of the above circles, add one drop of the appropriate antigen, which gives an 1030 Brussels
+91-44-22541131 Belgium
agglutination in the Screening Slide Test. Tel: +(32)2 732-59-54
E-mail:mail@[Link]
c) Mix the contents of each circle separately and spread it in the entire circle. +91-44-22541131
d) Rock the slide gently for one minute & observe for agglutination.