Culturing with Automated Culture Methods

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Culturing with Automated Culture Methods

• Hanh M. Khuu, M.D.
• February 26, 2004

Bacterial Screening in Platelets A Review of
Community Practices
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LR APL
AABB AABB AABB
FDA FDA FDA
HPC
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• Validation
• Written plan design, procedures, records, review
• Organism culture, dilution, colony count
• Seeding culture bottles
• Tracking bottles
• Confirmation of positive cultures

Process Validation
Proof of concept Validation for Intended
Use Process Validation
Validation for Intended Use 3 organisms 1 dilution
Instrument Validation 3 organisms 1 dilution
Instrument Validation 3 organisms 1 dilution
Instrument Validation 5 organisms 1 dilution
HPC
LR APL
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Study Design and Goals
Parallel Study CFR/USP vs.
BacT/Alert To compare the use of BacT/Alert
and the CFR/USP methods for sterility testing
of in-process and final product samples of cell
therapy products

• Seeded Study
• CFR/USP
• vs. BacT/Alert
• vs. Bactec
• To demonstrate that automated
• blood culture systems
• (BacT/Alert and Bactec) are
• at least equivalent to the
• CFR/USP method for
• sterility testing of
• cell therapy products

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Methods - Seeded Study

• 6 media preparations
• 3 culture systems (Bactec, BacT/ALERT, CFR)
• Aerobic and anaerobic media of each system
• 10 organisms
• 2 dilutions (10 CFU and 50 CFU)
• 3 replicates/organism
• 14 days incubation
• 0.5 ml sample

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Results Seeded Study
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Results Seeded Study
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Parallel Study

• November 1, 2002 November 30, 2003
• Two culture systems (BacT/Alert and CFR)
• A total of four bottles are used to culture each
  product
• Assessed twice per week
• 0.5 ml inoculum/bottle

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Parallel Study Results
Minimally manipulated
Minimally manipulated Selection Elutriation
2-20 days
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Questions??