Task | Identify relevant consensus test parameters | Screen candidate culture media with regard to mycoplasma cell growth and confirm the ability of the mycoplasma cells to penetrate a randomly chosen 0.2 μm rated penetration control filter | Choose challenge fluid by Task Force Committee member consensus Evaluate five candidate culture media with regard to mycoplasma cell growth and ability to penetrate a randomly chosen 0.2 μm rated penetration control filter in 5 different laboratories Choose culture media
| Confirm that the chosen media will generate test cells that will penetrate three different 0.2 μm rated filters, thereby ensuring a robust test of a 0.1 μm rated filter. When testing a 0.1 μm rated filter, the 0.2 μm rated filter will serve as a penetration control, similar to the role of a 0.45 μm rated penetration control filter when testing a 0.2 μm rated filter. |
Supporting Activity | Identify what test parameters will require testing to reach a consensus. | Testing was performed in a single independent (volunteer) laboratory. | Testing was performed in five different filter manufacturer laboratories. | Testing was performed in a single contract laboratory. |
Final Outcome | Consensus Test parameters to be evaluated:The test organism Challenge level The test pressure The challenge volume for a 47 mm disc The challenge fluid (selected in Phase 2) The positive control (penetration control): penetration of a 0.2 μm rated filter Culture media for the generation of test cells (this test parameter required testing to reach a consensus)
| All candidate media, except one, supported good growth of A. laidlawii and generated cells capable of penetrating a 0.2 μm rated penetration control filter. | A challenge fluid was chosen by consensus of the Task Force Committee: phosphate buffer A culture media was chosen (designated Mycoplasma Task Force Media) as a result of the testing and consists of20 g/L mycoplasma broth base (Beef heart infusion broth) 25 g/L yeast extract 100 mL/L of heat-inactivated horse serum (held at 55 °C for a minimum of 30 min)
| The test cells penetrated all three different 0.2 μm rated penetration control filters. Summary of final consensus test parameters:The test organism: A. laidlawii ATCC® 23206™ (NC 10116; Strain PG8) Challenge level (≥1 × 107 CFU/cm2 of test filter area) The test pressure: 30 psid (207 kPa) The challenge volume for a 47 mm disc: 200 mL The challenge fluid: phosphate buffer The positive control (penetration control): penetration of a 0.2 μm rated filter Culture media: Mycoplasma Task Force Media
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