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STANDARD OPERATING PROCEDURE

EYE CULTURE

1. Principle

Conjunctival swabs are submitted to the laboratory for detection of the bacterial agents that
may cause conjunctivitis, or inflammation and infection of the conjunctiva.

2. Materials

a. Gram smear: clean glass slide, gram smear reagents


b. Media - Blood Agar (BAP) and Chocolate Agar (CHOC), Mueller-Hinton agar

c. Incubator, 35-37°C with 5% CO2 (or a candle jar)


d. Inoculating loops

e. Reagents and discs required for organism identification and susceptibility testing

3. Specimen

It is preferable that both eyes be swabbed, even if the infection is unilateral. Swabs should
be collected prior to the instillation of topical antibiotics, and sent in transport medium.

4. Quality Control (QC)

Check that the patient name and identifiers on the specimen match that on the
accompanying requisition.

Ensure that all media and supplies used have passed the required QC and are used within
their expiry date.

5. Safety Precautions

Standard safety precautions for handling of patient specimens must be applied when
processing these specimens:

6. Procedure for Processing of Specimens

Direct smear: Use a clean dry glass slide to prepare the smear. Allow to dry, fix, and stain.

Examine the slide under the microscope; quantitate polymorphonuclear leukocytes (PMNs),
squamous epithelial cells and organisms seen.

Culture: Note that the culture plates are inoculated prior to the preparation of the smear.

3.6.11 SOP: Eye Cultures Page 1 of 2


STANDARD OPERATING PROCEDURE

Rub the swab over a portion of the plates and then streak the inoculums in 4 quadrants to
obtain isolated colonies. Incubate the plates in 5% CO2.

7. Interpretation

Examine the BAP and CHOC plates after 24 and 48 hours incubation.

Potential pathogens: S. aureus, H. influenzae, M. catarrhalis, N.gonorrheae, Group A Strep,


S. pneumoniae, Moraxella species, and P. aeruginosa.

For other organisms, a significant result is determined by the isolation of a moderate or


heavy growth of a potential pathogen correlated with the predominant organism on the
Gram stain. There should be >1+ pus cells on the Gram stain.

Full identification is required for all significant organisms

8. Reporting

a) Gram stain: Report with quantitation the presence of pus cells and organisms.

b) Culture:

Negative Report: "Commensal flora" or "No growth".

Positive Report: Quantitate all significant isolates and report with appropriate antimicrobial
susceptibilities.

If commensal flora is also present, report with quantitation.

9. Procedural Notes

The conjunctiva is colonized with normal flora such as coagulase negative staphylococci,
diphtheroids, alpha-hemolytic streptococci and Neisseria sp. The isolation of these
organisms is generally considered to be contamination. When these organisms are isolated
in the presence of pus, correlate with clinical findings in determining the work-up.

10. References

Clinical Microbiology Procedures Handbook, 3rd Edition, 2010. ASM Press, Washington DC.

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