Table 3. Mycoplasma detection methods (Also refer to the studies by Uphoff et al (1992) and Tuly and Razin (1996)).

Method	Sensitivity	Specificity	Advantages	Disadvantages
Microbiological culture (selective broth and agar).	High	High	European Pharmacopeia recommended test. Gives a definitive result.	Requires specialist microbiology lab. Relatively slow. Potential source of cross-contamination. Some strains are non-culturable.
Direct DNA stain, e.g. Hoechst 33 258 or DAPI	Low	Low (non-specific cytoplasmic DNA stains)	European Pharmacopeia recommended test. Rapid and cheap.	Reading and interpretation of test can be difficult and subjective.
Indirect DNA stain, (using indicator cells e.g. 3T3 or 3T6)	High	Low (non-specific cytoplasmic DNA stains)	Amplifies contaminant so easier to interpret than direct stain.	Indirect therefore slower and more time consuming than direct stain.
PCR, e.g., Universal Mycoplasma Kit1, Mycoplasma Plus2, MycoSensor2.	High	Medium (will not detect all Mycoplasma species)	Rapid and very sensitive. Several good commercial kits available.	False positives due to carry over contamination from positive controls and samples. False negatives due to PCR inhibitors in sample.
Enzyme Linked Immunosorbant Assay (ELISA), e.g. MycoProbe3, Mycoplasma PCR ELISA4.	Medium (High if amplified ELISA)	Medium (will not detect all Mycoplasma species)	Rapid and cheap. Useful and simple for screening large numbers of samples.	Amplified ELISAs have additional stages and are slower. Requires access to ELISA spectrophotometer.
Biochemical Detection, e.g. MycoAlert5.	Medium	High	Very rapid. Good for urgent testing of small sample numbers.	Requires access to a luminometer.

¹ ATCC (www.atcc.org).

² Agilent Technologies (www.genomics.agilent.com).

³ R&D Systems (www.RnDSystems.com).

⁴ Roche (www.roche-applied-science.com).

⁵ Lonza (www.lonza.com).

Examples of commercial mycoplasma testing kits are provided for information only with no endorsement intended or implied.