Detecting antimicrobial resistance: Difference between revisions

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= Phenotypic methods =
= Detecting antimicrobial resistance =
 
   
== Phenotypic methods ==
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== Broth dilution tests ==
 
=== Broth dilution tests ===
 
   
 
* Gold standard
 
* Gold standard
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** Trailing can occur; the MIC is read at the first spot where trailing begins
 
** Trailing can occur; the MIC is read at the first spot where trailing begins
   
=== Agar dilution ===
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== Agar dilution ==
   
 
* Concentration of antibiotic is incorporated into the agar
 
* Concentration of antibiotic is incorporated into the agar
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** Final inoculum of 10^4^ CFU per spot
 
** Final inoculum of 10^4^ CFU per spot
   
=== Disk diffusion ===
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== Disk diffusion ==
   
 
* Drug released from disk with an antimicrobial agent diffuses through the agar
 
* Drug released from disk with an antimicrobial agent diffuses through the agar
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* If on blood agar, must be read from the front
 
* If on blood agar, must be read from the front
   
=== Other ===
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== Other ==
   
 
* Potentiation using standard methods
 
* Potentiation using standard methods
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* Lateral flow assay
 
* Lateral flow assay
   
== Genotypic methods ==
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= Genotypic methods =
   
 
* Detected resistance may not predict antibiotic failure if genes are non-functional or not expressed, or may be expressed in insignificant levels
 
* Detected resistance may not predict antibiotic failure if genes are non-functional or not expressed, or may be expressed in insignificant levels
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** Sequencing
 
** Sequencing
   
== Other methods ==
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= Other methods =
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== MALDI-ToF ==
   
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[[Category:Antimicrobials]]
=== MALDI-ToF ===
 
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[[Category:Microbiology]]

Revision as of 20:12, 14 August 2019

Phenotypic methods

Broth dilution tests

  • Gold standard
  • Uses serial two-fold dilutions of the antimicrobial agent
  • Macrobroth has a final volume of 1-2 mL
  • Microbroth has a final volume of 50 µL
  • Determines an MIC and MBC (min. bactericidal concentration)
    • Read at lowest concentration that inhibits growth (i.e. no obvious growth)
    • Trailing can occur; the MIC is read at the first spot where trailing begins

Agar dilution

  • Concentration of antibiotic is incorporated into the agar
  • Each agar plate has a different concentration
  • Multiple inocula are set up on each plate
    • Final inoculum of 10^4^ CFU per spot

Disk diffusion

  • Drug released from disk with an antimicrobial agent diffuses through the agar
  • Zone diameter is inversely proportional to the log of the MIC
  • Interpretation usually uses reflected light reading above a black, nonreflective background; measured using a ruler on the back of an inverted dish
  • If on blood agar, must be read from the front

Other

  • Potentiation using standard methods
  • Colorimetric methods
  • Lateral flow assay

Genotypic methods

  • Detected resistance may not predict antibiotic failure if genes are non-functional or not expressed, or may be expressed in insignificant levels
  • Similarly, a negative results may not indicate susceptibility given genetic variation, novel mechanisms of resistance, or inhibition of amplification
  • Genotypic methods
    • PCR
    • Sequencing

Other methods

MALDI-ToF