LABORATORY DIAGNOSIS OF WOUND INFECTION

A. Clinical Presentation
Wound is a breach in the skin and the exposure of subcutaneous tissue
following loss of skin integrity caused by trauma, surgery, burns, ulcers,
fistulas, sinuses etc.
Wound infection can be either community- acquired or nosocomial in origin.
The presence of bacteria in a wound may result in:
contamination the bacteria do not increase in number or cause clinical
problems
colonisation the bacteria multiply, but tissues are not damaged
infection the bacteria multiply, healing is disrupted and wound tissues are
damaged (local infection). Bacteria may produce problems nearby (spreading
infection) or cause systemic illness (systemic infection)

B. Organisms Responsible
1) Staphylococcus aureus

7) Candida sp.

2) Coagulse Negative

8) Bacteroides sp.

Staphylococcus (CONS)

9) Actinomycetes sp.

3) Streptococcus pyogenes

10) Nocardia sp.

4) Enterococcus sp.

11) Prevotella sp

5) Enterobacteriaceae

12) Porhyromonas sp.

members
6) Pseudomonas sp.

13) Fusobacterium sp.

14) Clostridium sp.

C. Sample Collection & Transport

Incision and drainage

Wound swabbing and

Needle aspiration

Wound biopsy.

Blood for culture may be positive, in case of systemic spread of infection.

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Note:
1. Wound swab is most widely used, but may give misleading culture results by
detecting colonizing flora, rather than deep- seated pathogens.
2. Wound biopsy provides the most relevant information regarding the identity
of the involved pathogens, but is invasive and is reserved for wounds that are
failing to heal despite treatment.
3. Otherwise, aspirated pus from the wound is the preferred sample.

D. Macroscopic Examination
Examine the sample for colour, presence of blood, presence of granules
(typical of Actinomycetes, Nocardia)

E. Microscopic Examination
Gram stain: Gram- stained smear is examined for obtaining a preliminary
idea of the organisms involved, till the culture reports are available. Gram stain
reveals the presene of pus cells, bacteria and yeast cells.
F. Culture
1] Pus/Exudate: Inoculated in Blood agar, MacConkey agar and
Thioglycollate broth/RCM
Sabourauds Dextrose Agar may be inoculated if Candida is suspected.
2] Blood: Primary inoculation in BHI BiPhasic medium and subculture on
Blood agar and MacConkey agar.
G. Identification Tests
Based on the colony morphology observed on culture media, the following
biochemical tests may be performed for identification of the isolate:
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S. aureus or CONS: Catalase/Coagulase/Mannitol Fermentation

S. pyogenes: Catalase/Bacitracin Sensitivity/ PYRase

Enterococci: Catalase/Bile Aesculin/ PYRase/ Salt tolerance test/ Growth

at 100C; 450C; pH 9.6

Pseudomonas sp: Catalase/Oxidase/Oxidation- Fermentation/Pigment

Enterobacteriaceae:

Catalase/oxidase/IMViC/

TSI/Sugar

fermentation/

Nitrate Reduction/Urease/ PPA

Candida sp: Germ tube test/ Sugar Assimilation & Fermentation test/ Corn
Meal Agar

H. Antibiotic susceptibility testing

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