Disk Diffusion Test Flashcards
Test for S. pneumoniae’s penicillin resistance
Oxacillin disk screen
Medium for Oxacillin disk screen
MHA with 5% sheep blood and 1 µg oxacillin
Incubation for Oxacillin disk screen
35°C, 5-7% CO2, 20-24 hours
Result interpretation for Oxacillin disk screen
≤21 mm = Resistant, intermediate, or susceptible (requires MIC testing)
Test for MRSA to penicillinase-resistant penicillins
Oxacillin screen agar
Medium for Oxacillin screen agar
MHA with 6 µg oxacillin and 4% NaCl
Incubation for Oxacillin screen agar
30-35°C, 24 hours
Positive result for Oxacillin screen agar
Growth = Resistance
Negative result for Oxacillin screen agar
No growth = Susceptible
Test to detect mecA-mediated oxacillin resistance
Cefoxitin disk diffusion
Medium for Cefoxitin disk diffusion
MHA with 30 µg cefoxitin
Incubation for S. aureus and S. lugdunensis
33-35°C, 16-18 hours
Incubation for CoNS
35°C, 24 hours
Result for S. aureus and S. lugdunensis
≤21 mm = mecA positive; ≥22 mm = mecA negative
Result for CoNS
≤24 mm = mecA positive; ≥25 mm = mecA negative
Test for Vancomycin resistance in VRE and VRSA
Vancomycin screen agar
Medium for Vancomycin screen agar
BHI with 6 µg vancomycin/mL
Incubation for Vancomycin screen agar
35°C, 24 hours
Positive result for Vancomycin screen agar
Growth = Resistance
Negative result for Vancomycin screen agar
No growth = Susceptible
Purpose of β-lactamase detection
Detects β-lactamase production, resistance to Ampicillin, Penicillin, and derivatives (-cillin)
Organisms tested for β-lactamase production
Enterococcus, N. gonorrhoeae, H. influenzae, Bacteroides
Method for Chromogenic Cephalosporin test
Nitrocefin disk + saline + bacterial loop
Incubation for Chromogenic Cephalosporin test
22-25°C, 10 minutes (Staph. 60 minutes)
Positive result for Chromogenic Cephalosporin test
Red/Pink = β-lactamase producer
Method for Acidimetric β-lactamase detection
Penicillin + phenol red indicator + bacteria
Incubation for Acidimetric β-lactamase detection
22-25°C, 10-15 minutes (Staph. 60 minutes)