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Microbial Limit Testing Technology Overview

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Counts only viable organisms. DISADVANTAGES. Incubation time (48-72 hours) ... Microbial Viability. Only interested in viable microorganisms ' ... – PowerPoint PPT presentation

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Title: Microbial Limit Testing Technology Overview


1
Microbial Limit TestingTechnology Overview
  • Advisory Committee
  • for
  • Pharmaceutical Science
  • July 19, 2001

2
Compendial MethodsUSP
  • Plate Counts- Colony Forming Units
  • Growth of microorganisms on solid medium
  • Count visible colonies
  • MPN- Most Probable Number
  • Multiple serial dilutions of sample
  • Based on statistical distribution of organisms in
    a sample

3
Compendial Methods
  • ADVANTAGES
  • Simple
  • Counts only viable organisms
  • DISADVANTAGES
  • Incubation time (48-72 hours)
  • Not all organisms will grow on a single medium

4
Microbial Viability
  • Only interested in viable microorganisms
  • New methods must only count viable organisms
  • Viability indicators needed

5
Esterase DetectionChemScan, Chemunex
  • Enzyme ubiquitous in microorganisms
  • Chem-Chrome, a non-fluorescent fluorescein
    derivative cleaved by esterase to fluorescein

6
ChemScan Procedure
  • Filter Sample
  • Expose membrane to reagent
  • Membrane analyzed by laser-scanning

7
ATP BioluminescenceMicroStar(TM)
MicroCount(TM), Millipore
  • ATP- Primary energy source for all organisms
  • Luciferin/luciferase (reagent) reacts with ATP to
    produce light

8
MicroStar(TM) Procedure
  • Filter Sample
  • Place membrane on solid medium for brief
    incubation
  • Disrupt cells to release ATP
  • Add bioluminescence reagent to membrane
  • Detect light with CCD and computer analysis
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