Mycoplasma Detection Methods: Culture Isolation
Often regarded as the gold standard method for Mycoplasma detection, the culture isolation detection method uses a combination of selective growth media and incubation conditions to positively enrich for any Mycoplasma present in a sample. Most Mycoplasma contaminants can be detected by growth on standardised agar, with the exception of certain strains of M.hyorhinis.
Colonies of Mycoplasma exhibit a distinctive "fried egg" morphology (see image) when viewed under a plate microscope. The assay requires a 28 day test interval before a definitive result can be obtained.
Colony of Acholeplasma laidlawii showing typical "fried egg" morphology.
Viewed under magnification
ECACC offer the culture isolation method to both regulatory protocols and in-house variations of pharmacoepial methods.
Mycoplasma Detection Methods:
| Assay type | Assay Characteristics | ||||
|---|---|---|---|---|---|
| Sensitivity | Speed | Species Range | ECACC In-house Protocol |
US FDA Approved Protocol |
|
| PCR | Low/Medium | 1 day | Majority | Yes | No |
| DNA Stain | Medium | 2-3 days | All | Yes | Yes* |
| Culture Isolation | High | 3-4 weeks | Majority | Yes | Yes |
*European Pharmacopoeia protocol also available by arrangement