Expt.

8 ◦ Serum Agglutination in slide are approximately equivalent to

Febrile Agglutination Tests the test tube dilutions
◦ For the detection of antibodies against certain bacteria. Serum Volume Approximate Tube Dilution
◦ Diagnose bacterial infections associated with persistent 0.08 ml 1:20
pyrexia(fever) 0.04 ml 1:40
─ Typhoid fever(Salmonella)
0.02 ml 1:80
─ Typhus fever(Ricketssia)
0.01 ml 1:160
─ Brucellosis(Brucella)
─ Tularemia(Francisella) 0.005ml 1:320
Typhoid Fever ◦ Interpretation
◦ Transmitted through ingestion of contaminated food or food ─ Score of (2+) or greater is considered a positive test
products ─ Significant titer is 1/80
◦ Gastroenteritis ─ High or rising titer of “O” antigen = ACTIVE INFECTION
◦ Bacteremia and Extraintestinal Infections ─ High or rising titer of “H” Antigen= Past
◦ Enteric Fever infection,convalescence, past immunization,exposure to
Typhoid Fever Antigens area where there is a prevalence of typhoid fever
◦ H antigen (Hauch) ─ High titers are observed on all antigens= Mixed Infections
─ Occur in flagella
─ Thermolabile Rickettsial Infections
─ Specific for the given species ◦ Results from either an arthropod bite or contamination with
─ Detect Flagellar agglutinins its feces
─ Preparation: add 2% formalin ◦ Vasculitis
◦ O Antigen(Ohne) ◦ Cell necrosis
─ Directly associated with bacterial body ◦ Thrombosis of blood vessels
─ Thermostable ◦ Skin rashes
─ Non specie specific ◦ Organ dysfunction
─ Detect somatic agglutinins against Salmonella and Proteus
(OX2, OX19,OXK) ◦ Weil Felix Test
─ Preparation: add 5%phenol ─ Non specific rickettsial test based on cross reacting
◦ K antigen(kapsel) antibodies
─ Capsules or as envelope surrounding the bacterial body ─ Antibodies of typhus fever cross react with polysaccharide
─ Vi ( Variety of K Ag) – typhoid carriers O antigen of some Proteus species
◦ Widal Test ◦ Antigens used:
─ Measures the presence of anti-O and anti-H antibodies ─ Proteus OX2 and Proteus OX19: obtained from Proteus
using bacterial suspensions of killed S. typhi and S. vulgaris
paratyphi. ─ Proteus OXK: obtained from Proteus mirabilis
─ Slide Method: Screening ◦ Weil Felix Test Procedure
─ Tube Method: Confirmatory 1. Using 0.1 ml serological pipette, deliver 0.08ml,
◦ Widal Test Procedure 0.04ml,0,02ml, 0.01ml and 0.05ml quantities of serum to
1. Using 0.1 ml serological pipette, deliver 0.08ml, the circles of the slide from left to right.
0.04ml,0,02ml, 0.01ml and 0.05ml quantities of serum to the 2. Add one drop of antigen suspension just below each
circles of the slide from left to right. quantity of serum.
2. Add one drop of antigen suspension just below each quantity 3. Mix using different applicator sticks
of serum. 4. Gently tilt slide back and forth for not more than one
3. Mix using different applicator sticks minute.
4. Gently tilt slide back and forth for not more than one minute. 5. Observe for agglutination
5. Observe for agglutination ◦ Manner of Reporting Results (same with Widal Test)
◦ Manner of Reporting Results ◦ Interpretation
─ Score of (2+) or greater is considered a positive test
Manner of Appearance of test result ─ Significant titer is 1/160
Reporting
Reaction with Proteus
Negative No agglutination is observed Strain
OX2 OX19 OXK
Trace Less than 25% of the organisms are agglutinated 1. Rocky Mountain Spotted +1 +4 -
Fever(R.ricketssi)
1+ 25% of the organisms are agglutinated 2.Murine Typhus(R.mooseri) +1 +4 -
2+ 50% of the organisms are agglutinated 3.Epidemic Typhus(R.prowazeki) +1 +4 -
4.Brill’s Disease +1 +4 -
3+ 75% of the organisms are agglutinated 5.Scrub Typhus(R.tsutsugamushi) - - +3

4+ 100% of the organisms are agglutinated

GRANULAR TYPE of Agglutination(Fine Aggregates):
─ Typhoid O, proteus, Brucella, Francisella antigens
FLAGELLAR TYPE of Agglutination(Loose or Fluffy clumps):
─ Typhoid H, Paratyphoid A, Paratyphoid B