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)
Positive result for Acidimetric β-lactamase detection
Yellow = β-lactamase producer
Method for Iodometric β-lactamase detection
Penicillin + iodine-starch complex + bacteria
Incubation for Iodometric β-lactamase detection
22-25°C, 10 minutes (Staph. 60 minutes)
Positive result for Iodometric β-lactamase detection
Colorless/White = β-lactamase producer
Purpose of ESBL detection
Detects resistance to Penicillin, Monobactams, and Cephalosporins
Organisms tested for ESBL
Klebsiella, E. coli
Method for ESBL detection
Disk diffusion with Cefotaxime (CTX), Ceftazidime (CAZ), and Clavulanic acid (CLA)
Incubation for ESBL detection
35°C, 24 hours
Positive result for ESBL detection
Larger ZOI with CTX/CAZ + CLA compared to CTX or CAZ alone
Purpose of Aminoglycoside screen
Detect high-level resistance to Aminoglycosides in Enterococci
Medium for Aminoglycoside screen
BHI broth with Gentamicin and Streptomycin
Incubation for Aminoglycoside screen
35°C, 24 hours
Positive result for Aminoglycoside screen
Growth = Resistance
Negative result for Aminoglycoside screen
No growth = Susceptible
Purpose of ICR test
Detects inducible clindamycin resistance
Organisms tested for ICR
S. aureus, S. lugdunensis, S. pneumoniae, beta-hemolytic Streptococci
Media for ICR test
5% Sheep BAP or MHA with 15 µg Erythromycin (E) and 2 µg Clindamycin (Cd)
Incubation for ICR test
35°C, 16-18 hours (for S. aureus, S. lugdunensis), or 35°C, 20-24 hours with 5% CO2 (for S. pneumoniae, beta-hemolytic Streptococci)
Positive result for ICR test
Flattening of CD zone (D-zone) = Clindamycin resistant
Purpose of Modified Hodge test
Detects carbapenemase production in Enterobacterales
Media for Modified Hodge test
MHA
Disk for Modified Hodge test
10 µg Meropenem or Ertapenem
Incubation for Modified Hodge test
35°C, 16-18 hours
Positive result for Modified Hodge test
Clover-leaf like appearance, indented or enhanced growth
Purpose of CNP test
Confirmatory test for carbapenemase production
Reagents for CNP test
Imipenem, Phenol red indicator
Incubation for CNP test
35°C, 2 hours
Positive result for CNP test
Tube A = Red/Orange, Tube B = Yellow
Determines MIC of 1 antimicrobial against 1 bacteria
Gradient diffusion testing
1 gradient of antibiotic concentration in a plastic strip
E-test/Epsilometer
1 gradient of antibiotic concentration in a paper strip
Liofilchem
Concentration causing 99.9% reduction in CFU/mL after broth dilution and subculture
Minimum bactericidal concentration (MBC)
Antibiotic inhibits but does not kill bacteria, linked to beta-lactam resistance
Tolerance
Serum bactericidal test using patient’s own serum
Schlichter test
Highest drug concentration in the serum
Peak specimen
Lowest drug concentration in the serum
Trough specimen
Drug peak concentration collection: IV 30–60 mins, IM 60 mins, Oral 90 mins after admin
Timing of specimen collection
Bacteria inoculated in antimicrobial concentrations, incubated, and colony counts measured over time
Time-kill assay
Effectiveness of multiple antimicrobials on a single bacterial strain
Synergy test
Normal flora of the human microbiome
Colonizer
Bacteria that multiply on the host
Resident
Bacteria that do not multiply on the host
Transient