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LABORATORY DIAGNOSIS OF BACILLARY DYSENTERY

A. Clinical Presentation
Short incubation period (1-7 days, usually 48 hrs)
Frequent passage of loose, scanty faeces containing blood & mucus
Abdominal cramps & tenesmus
Fever & vomiting may be present
Complications include arthritis, toxic neuritis, conjunctivitis & in
children, intussusceptions
Hemolytic uremic syndrome occur as complication in severe cases
B. Organisms responsible
Shigella sp: 1. S. dysenteriae (Group A)
2. S. flexneri (Group B)
3. S. boydii (Group C)
4. S. sonnei (Group D)
C. Specimen collection & Transport
Sample
Fresh stool (specimen of choice)
Mucus flecks
Rectal swabs
Sample Collection
25 ml screw-capped, wide-mouthed plastic container.
1 teaspoon (5 mL) of faeces is collected into the container.
Transport of Specimen
Transmit the specimen as soon as possible.
If delay is unavoidable, transport medium- Buffered glycerol saline is
used.
D. Macroscopic Examination
1. Characterized by the presence of mucus & blood
E. Microscopic Examination (Wet mount)
A drop of fresh stool sample is placed onto a drop of Methylene Blue
taken on a slide.
A coverslip is applied & examined under high dry objective lens (40x).
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Observations
Presence of faecal leucocytes
Presence of red blood cells
F. Culture: Inoculated in solid selective media & enrichment broth
Solid selective Media
Media
MacConkey agar

Colony characterstics
Pale in color except S.sonnei which is a late
lactose

fermenter,

become

pink

when

incubation is prolonged beyond 24 hrs.


Xylose Lysine

Red in color without black center

Deoxycholate agar
Salmonella Shigella agar

Red in color without black centre

Note: Do not use SS agar as it frequently inhibits the growth of S. dysenteriae


serotype 1.
Enrichment broth
Selenite F broth
After 6 hrs of enrichment at 37C, subcultures are done on selective medium
such as XLD agar
G. Identification Tests (Biochemical tests & Slide Agglutination)
Tests
Motility

Result
Non-motile

Oxidase

Negative

Nitrate reduction test


Sugar fermentation test

Positive
Glucose fermented with acid no gas
Lactose not fermented except S. sonnei
Mannitol fermented with acid no gas except S.
dysenteriae
Maltose not fermented
Sucrose not fermented except S.sonnei
Xylose not fermented
Positive
Negative
Positive
Negative
Negative

Catalase
Indole production
Methyl red
Voges-Proskauer
Citrate Utilization test

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Urease
Triple Sugar Iron agar

Negative
Alkaline slant / Acidic butt ( K/A, no gas, no
H2S)
Phenylalanine deaminase Negative

Slide Agglutination Test


Serotyping for Shigella to be guided by biochemical reactions
Mannitol Fermentation

Not Fermented
(S.dysenteriae)

Fermented with acid, no gas

Lactose Fermentation
For Confirmation by Serotyping

Test S.dysenteriae polyvalent antisera

If Positive
Test S.dysenteriae monovalent antisera
Non Lactose Fermenter

Late Lactose Fermenter

(S. flexneri, S.boydii)

(S.sonnei)
For Confirmation
Test S.sonnei antisera

For Confirmation
Test S. flexneri polyvalent antisera

If Positive
Test S. flexneri monovalent antisera

If Negative
Test S.boydii polyvalent antisera

If Positive
Test S.boydii monovalent antisera
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