Corneal transplantation, or keratoplasty, is a common procedure aimed at restoring vision in patients with corneal diseases.
The success of this surgery heavily relies on the quality of the donor cornea, which must be free from infectious pathogens. To ensure the safety and effectiveness of corneal grafts, culture and sensitivity testing of corneal donor rims is crucial.
1] Sample Collection -
A] Aseptic Technique: Strict aseptic protocols are followed to avoid contamination. Surgical instruments and collection containers must be sterile.
B] Sample Preparation: The donor rim is carefully dissected from the rest of the corneal tissue and placed in a sterile container for transport to the microbiology lab.
2] Culture Techniques -
A] Inoculation on Culture Media: The corneal rim is inoculated onto various culture media designed to support the growth of bacteria and fungi. Common media include blood agar, chocolate agar, and Sabouraud dextrose agar for fungi.
B] Incubation: The inoculated media are incubated under controlled conditions, typically at 35-37°C, for a duration that may range from 24 to 48 hours. Fungal cultures may require longer incubation periods.
3] Identification of Microorganisms -
A] Colony Morphology: The appearance of colonies is noted, as different organisms exhibit unique characteristics.
B] Biochemical Tests: Specific tests (e.g., catalase, oxidase) are conducted to identify the microorganisms.
C] Molecular Techniques: Advanced molecular methods, such as polymerase chain reaction (PCR), may be utilized for rapid and accurate identification of pathogens.
4] Sensitivity Testing -
A] Disk Diffusion Method: This method involves placing antibiotic-impregnated disks on agar plates inoculated with the identified microorganisms. The zones of inhibition are measured to evaluate susceptibility.
B] Broth Dilution Method: This technique determines the minimum inhibitory concentration (MIC) of antibiotics, indicating the lowest concentration needed to inhibit bacterial growth.
1. Susceptibility Results -
A] Susceptible: Indicates that the organism is likely to be effectively treated with the antibiotic at achievable serum concentrations.
B] Intermediate: Suggests that the antibiotic may be effective but could require higher doses or may not be effective in some patients.
C] Resistant: Indicates that the organism is not likely to be affected by the antibiotic, guiding the clinician away from using it.
2. Clinical Implications - The results of culture and sensitivity testing will inform the choice of prophylactic antibiotics for the recipient of the corneal transplant. For instance, if the testing identifies Pseudomonas aeruginosa as a potential pathogen, the clinician may opt for specific antibiotics known to be effective against it.
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1. Culture Method
2. Sample
3. Colony Count
4. Organism(s) Isolated
5. Culture Report: Culture yields growth of
6. Culture isolated after 7 days :
7. Culture isolated after 14 days:
8. Culture isolated after 21 days:
9. Ampicillin
10. Amikacin
11. Amoxicillin clavulanate
12. cefoperazon+sulbactam
13. Cefuroxime
14. Cefepime
15. Cefotaxime
16. Ciprofloxacin
17. Ertapenem
18. Gentamicin
19. Imipenem
20. Meropenem
21. Norfloxacin
22. Nitrofurantoin
23. Piperacillin-tazobactam
24. Trimethoprim-Sulfamethoxazole (Cotrimoxazole)
This testing is crucial to prevent postoperative infections, ensuring that the donor cornea is free from pathogens that could compromise the recipient's vision and overall outcome of the transplant.
Risks include infection, rejection of the graft, and complications related to the surgery itself. Ensuring the safety of the donor cornea through culture and sensitivity testing helps mitigate these risks.
Various culture media may be used, including blood agar, chocolate agar, and Sabouraud dextrose agar for fungi. The choice of media depends on the suspected pathogens and their growth requirements.
