ACCUCARE
WIDAL TUBE TESTS
ORDER INFORMATION PROCEDURE FOR TUBE METHOD:
Kit Size Cat. No. 1. Take 4 sets of 8 clean dry test tubes (10x75 mm) for “O”, “H”, A “H” &
B”H” antibody detection.
4 x 50 ml WIT 1
2. Dilute each serum sample as follows:
INTRODUCTION:
Test Tube 1 2 3 4 5 6 7 8
Serum Saline
Typhoid fever is an acute infectious disease characterized by definite 1:20 1:40 1:80 1:160 1:320 1:640 1:1280
Dilution Control
lesions in peyer’s patches, mesenteric glands and spleen accompanied Normal Saline 1.9ml 1ml 1ml 1ml 1ml 1ml 1ml 1ml
by fever, headache and abdominal symptoms. It is also called ‘enteric’
fever. Most frequent and dangerous complication in the late stages of Patient’s
this fever is intestinal haemorrhage and perforation. 0.1ml
Serum -- -- -- -- -- -- --
(undivided)
The causative organism of the typhoid fever is a gram negative bacillus,
Salmonella. In our country typhoid is commonly caused by Salmonella Transfer ➞ ➞ ➞ ➞ ➞ ➞
Diluted serum -- 1ml 1ml 1ml 1ml 1ml 1ml
typhl, S. Paratyphi A and S. Paratyphl B. These organisms possess
somatic (O) as well as flagellar ‘H’ antigens. The three serotypes have Appropriate
0.5 0.5 0.5 0.5 0.5 0.5 ml 0.5 ml 0.5 ml
common ‘O’ antigen but possess different ‘H’ antigens. During infection Antigen ml ml ml ml ml
with these bacteria, antibodies appear in the patient’s, within 2 to 5
weeks. A rising titre is highly significant for diagnosis of an active
infection. 3. Mix well and then incubate at 37°C for 16 to 20 hrs, and observe for
agglutination.
The detection of these antibodies in suspected patients forms the basis 4. ‘O’ antigen shows granular agglutination and ‘H’ antigen gives floccular
of the ‘Widal’ test. The antigens used in Widal Kit are standardized, appearance. Note the highest dilution showing clearly visible
smooth suspensions of killed bacilli, which are stained for their easy agglutination with naked eye.
identification in agglutination reactions. Two tests procedure are 5. saline control should remain unchanged which indicate a negative test.
employed. The tube agglutination test is done as a confirmatory test to
establish the presence or absence of an homologous antibody. INTERPRETATION OF RESULTS:
PRINCIPLE: Agglutination titre of 1:80 or more is significant. An increase in Titre, 4 to 5
days after the first test is suggestive of an active infection
When a patient’s serum containing antibodies to S. typhi and S.
paratyphi is mixed with the respective antigen, agglutination takes place NOTES:
which indicates the presence of antibodies to a particular antigen (O, H,
AH, or BH) 1. Individuals vaccinated with TAB vaccine may show moderate titre
against all three ‘H’ antigens.
SAMPLE COLLECTION AND PRESERVATION 2. Persons who suffered with enteric fever in the past may show moderate
Fresh serum samples should be used. titre when suffering from other unrelated illness. Such anamanesic
appearance of agglutination can be identified. If the patient is tested
REAGENT PREPARATION AND STORAGE again after few days. Only a rising titre is indicative of true infection.
3. The use of positive, negative and saline controls are recommended
The sample can be stored at 2-8°C if not immediately used. along with serum specimen.
REAGENT STABILITY BIBLIOGRAPHY
All reagents are stable upto the expiry date mentioned on the label
when stored at 2-8°C. Do not freeze. 1. Felix A, Brit Med. J. 11, 597 (1942)
2. Protel, R.L. et al, Lancet, 11.330 (1971)
3. Cruicks, R. Medical Microbiology.
12th Edition P. 403 (1982)
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