Salmonella typhi
simen Blood, faeces, urine
(Bacilli are excreted in faeces from initial part of the disease)
Iture Media Macconkey's Agar
lony Character On Macconkey's Agar colonies are Pale (NLF), circular, low convex
moist, smooth and translucent
ram Staining Gram Negative Non sporing Bacilli arranged haphazardly
lotility Hanging drop preparation shows organisms are motile.
(They have peritrichous flagella)
Biochemical Name of theTest Result
Reactions Indole Production Test Negative
Urease Test Negative
Citrate Uilization test Negative
Motile & Fermented
Manitol
Fermented but no Gas
Glucose Fermentation
Lactose Fermentation Not fermented
Further Tsts Of Name of the Test Result
Identification Catalase Positive
Oxidase Negative
Methyl Red Test Positive
VP Test Negative
Nitrate Reduction Test Positive
Serotyping Serotyping with specific antisera
can be done. The specific antisera
used are factor 9 (0) antisera and
polyvalent H antisera.
Final Diagnosis From the above parameters the organism is diagnosed as Salmonella typhi.
Note: Students have to draw the diagram as shown in the demonstration class.
Iutruluction
enteric lever.
done or sroloical diagnosis of
Iis
This is at lor measurement ofll & 0agglutinins for typhoid ¶typhoid bacili.
H&0antigen of S. typhi
& Hantigen of S.
The antigens usd ane
is not cmployed as they cross rcact
paratyphi
with typhoid A& 3. The
ary phoid () antigen O antigen duc
ofavommon lactor ((-12). to
I1, T0. A. Bl antigens are available
commercially. haring
Short, round botto lubes are used for lhe test.
These tubs are kept in a midal rack. A total Iof 4 rows consisling of 7 tubes in
nade. each row are
I Row: lor TH
a Rou:for TO
3 Row: for AH
4 Roy: for Bll
Procedure
Afirst 0.`cc of nommal saline is added to tube no. 2-7
in all
4.5cc nonnal saline is taken and 0.5 cc of test sera is added. the rows. Then in a separate
The dilution of tube
tuke is 1:10(N1aster dilution). test serum of this
Now. 0.5r from the master dilution is
transferred to tube I& tube
lest serum is l:10 in tube l and i:20 in tube2. 2.Therelore
Serial doubling dilution are no the dilu'ion of
no óby transferring 0.5cc of
give the dilution of test serummalerial by the help of a graduated pipetle from tube 2to upto (ute
made
the 7 tube thus contains only from tube l to tube 6| |:10. 1:20. 1:40. I:80. tube 6
0.5cc of normal saline. No test 1:14li, I:320] and
10 17 (0.5cc of test serun serum will be
from T6 is to be thrown off), translerred from Tó
The 7 ube will scrve as
Non 0.5cc of|l &O anigen contrd.
antigen is added to all the
marked tubes.
Alter addition of antigen to all
1:40. 1:80. 1:160. 1:320, 1:640.the tubes. the tinal dilution of test serun in Tl to
Now the widal rack is T6 are I:0.
Result: incubaled at 37°C overnight.
Hagglutiation leads
lo formation of
loose cotton nooly
O
aygluination is secen as a disc like clumps.
Amugnifying lens pattern at the bottom of
should be used for tubes.
Lnterpretalion: detecion of the small
granules.
Widal lest should be
interpreled laking ia lo accuunt
the followin
1|
Widal Test
patients serum in each Salmonclla suspension is rcad at the highest dilution
I)The titre of'
of l in 320.The titre will be 320.
enteric fever .So the ncgative resutt
The positive rcaction is first detected at the 7" to l0" day of3d 4 weck.A flcr 4h week the titre
stcadily till
inearly slage is not conclusive. The titre rises
gradually declines.
is
2)Demonstration of 4 fold rise of titre of antibody by testing 2 or more serum samples
in the discase .a rise may not be
more meaningful than a single test. Ifthe I sample is taken late
seen instead a fall inthe titre may be seen in some cascs.
because
3) For determining type of infection H reaction is more rcliable than O reaction
diflerent Salmonella have O antigen in common.
a
4)il positive result in a single test indicates the prescnce of enteric fever .Result of'
single test should be interpreted with caution. It is difficult lo luy down levcls of significance
though it is generally stated that titre of I00 or more for H agglutinins and 200 or more for o
agglutinins.It is necessary to obtain the distribution of agglutinin levels in nornmal sera in
different areas (i.e local titrre). In kolkata it is 80 in subclinical Salnmonclla infection
(Salmonellosis).
5) Hantibody persist for many months but "0" antibody tends to disuppwar sooner within
6 months,
6)Agglutinins may be prckil on account of prior discise. inapperant
infection in:munization. Persons who have received TAB vaccine will generally have antibody
against sal1monella typhi. Salmonella paratyphi A and B while in case of inteetious antibodies
nillbe scen ayuinst thc infectious species:
7)Persons who have had prior infection or immunization may give rise to an anamnestic
reaction during an unrelated fever. This may be dilferentiated from intections by repetition of
lest afier a week.The amnestic reaction will only show a transient rise while in enteric fever, the
rise is sustained. This is due to polyclonal activation of immunocompetent cells. As a result of
unrelated fever Salmonella antibody also rises transiently but due to lack of' repeated specilic
stimulisation the level falls quickly.
8) Bacterial suspension used as antigen should be free lrom fimbriae utherwise false
positive reaction will occur. Fimbrial antigen acts as nonspecilic antigen cnd react with the
antibody of infected individual.
9) Cases reated early with chloramphenicol may give pior agglutination response. Ir'
antibody is already present. there s ill be no rise in titre.
