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WIDAL TEST

Diagnostic Test for Typhoid Fever

TYPHOID FEVER

Mode of Transmission - ingestion of contaminated food or through contaminated hands Infections a.) Gastroenteritis b.)Bacteremia and Extraintestinal infections c.)Enteric Fever (Typhoid Fever)

PRINCIPLE

Human serum containing Anti-O and Anti-H antibodies , visibly agglutinate a suspension of Salmonella O and H antigens

Anti-O and Anti-H antibodies were measured using bacterial suspension of killed Salmonella typhi and Salmonella paratyphi

MATERIALS

Reagents -Serum -Salmonella O antigens (Groups A, B, C, D) -Salmonella H antigens (Groups a, b, c d) Glassware and Apparatus -Serological pipetes, 0.1 mL -porcelain spot plate -wooden applicator sticks

VOLUME OF SAMPLE

Patients Serum
0.08 mL 0.04 mL 0.02 mL 0.01 mL 0.005 mL

Dilution
1:20 1:40 1:80 1:160 1:320

PROCEDURE
Dispense the required Px serum on each of the well (Use 0.1mL serological pipette )

Add one drop of antigen suspension on the Px serum

Mix the serum and antigen using a wooden applicator stick

Tilt the spot plate back and forth and read within 2 minutes

Observe for agglutination using any good direct light

ANTIGEN

Thermostable somatic (Ohne or O antigen) Thermolabile flagellar (Hauch or H. Antigen)

Thermolabile capsular (Kapsel or K antigen)

Somatic ( O Ag)

- body of the organism


-used to detect somatic agglutinins against Salmonella - Salmonella O Ags (Group A, B, C, D) - preparation: extracting bacterial cultures using phenol or alcohol -reaction: fine, compact, not easily dispersed aggregates Flagellar (H Ag) - appendage of bacteria

- Salmonella H Ags (Group A, B, C, D)


- preparation: saline containing 2% formalin -reaction: loose, floccular, course, easily dispersed aggregates Capsular (K Ag) - capsule of bacteria -varieties of K Ag (B, L, Vi) - detection is carried out to confirm enteric infections

ANTIBODY
Anti-O

Anti-H
Anti-K

FEBRILE AGGLUTININS FOUND IN THE SERUM


O agglutinins - They are especially important because the titer of o agglutinins rises earlier in the disease and then drops faster than H agglutinnins. Oagglutinins are not effected by immunization. H agglutinins - They are usually unimportant because the H titer are slow to rise in a disease but remains elevated for several years. H titer increase after immunization.

MANNER OF REPORTING
100% of organism agglutinates = ++++ (+4)

75% of organism agglutinates = +++ (+3)


50% of organism agglutinates = ++ (+2)

25% of organism agglutinates = + (+1)


<25% of organism agglutinates = +- (trace) No agglutination = 0 (non reactive)
+2 is considered a positive test

INTERPRETATION
titer to O Ag = ACTIVE INFECTION

titer to H Ag = PAST INFECTION

titer to Vi Ag = CARRIER

titer to all Antigens = MIXED INFECTION

THANK YOU!!!
Roasa, Priscilla Grace Rubico, Faye Marjorie C. Salvador, Franz Raniel Sanchez, Charisse Santos, Samuel 3DMT