Anaerobic Bacteriology

Identification

June 2012

Identification can be further accomplished through the use of rapid commercial biochemical kits such as the Rapid ID-ANA II, API 20A, Vitek, and others.  All have variable accuracy.  Generally, they can identify to a species level approximately sixty percent of clinically significant anaerobes.  Again, one may choose to use PRAS biochemical’s but they are expensive, cumbersome, and generally, not necessary in the clinical laboratory.  Gas liquid chromatography for the detection of all fatty acids may also be employed.  Currently the new standard for identification and taxonomy of anaerobes is the use of 16S ribosomal RNA sequencing, which unfortunately is not available in many clinical laboratories.  In the future, MALDI-TOF mass spectrometry will probably be employed because it is such a rapid and simple methodology.  However, it is expensive equipment wise and at the present time because of limited database, it will not achieve complete identification of anaerobes.

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• Introduction
• Objectives
• Anaerobic Bacteria
• Principal Anaerobic Pathogens
• Specimen Selection: Avoid Contamination With Normal Flora
• Inappropriate Specimens
• Collection and Transport
• Anaerobic Transport Vials
• Anaerobic Transporters
• Anaerobic Culture Media
• Primary Culture CO₂ Holding Jar with Flow Meter
• Incubation
• Anaerobic Jars Set Up Using the Anaero-Pack
• Anaerobic Glove Box
• Anaerobe Culture Triage
• Anaerobe Culture Triage (cont.)
• Identification
• Colonial Morphology
• Pigmented Colonies
• Rapid Identification Using Gram Stain
• Identification
• 16S Ribosomal RNA Sequence of Bacteroides fragilis
• Antimicrobial Susceptibility Testing
• AST Methods
• Illustration of the Components of the E-test Method
• Susceptibility Testing by E-test
• Anaerobe Antimicrobial Panels
• References
• Special Thanks
• Questions