Mycoplasma Information

Mycoplasma Facts

• 35% of cell cultures worldwide are contaminated with mycoplasma¹, reaching 80% in some countries².
• Three major sources of mycoplasma: infected lab personnel, infected cells, and infected media.
• 80% of lab technician's throat swabs are mycoplasma positive³. Human-sourced species account for 50% of contaminated cultures¹.
• Sneeze, cough, and pipetting can transmit mycoplasma easily.
• Present on lab technician's hands and lab equipment.
• Survives on the surface of a laminar flow hood for 6 days.
• Does NOT cause visible changes in growth medium turbidity or pH (color).
• Does NOT kill the cells outright.
• Infected cells still look healthy with unapparent changes to growth rate and morphology, but hundreds of host protein expressions change and the NFkB signaling pathway is activated.
• Invisible under a microscope even at concentrations greater than 10⁷ copies/ml.
• Resistant to penicillin/streptomycin and other common antibiotics.
• Cannot be removed by 0.2µm filter "sterilization".
• A single contaminated culture can quickly infect other cultures in the laboratory.
• Once detected, the best solution is to discard the contaminated cells.

Good Habits to Minimize Mycoplasma Contamination

• Follow stringent aseptic culture techniques.
• Do NOT sneeze, cough, or talk loudly in the cell culture area.
• Sterilize the hood surface every time (70% ethanol).
• Always wear gloves.
• Work with only one cell culture at a time.
• Designate separate media and reagents for each individual cell line.
• Sterilize lab equipment (hemacytometer and disposal trays) routinely.
• Test for mycoplasma when thawing new cells.
• Test for mycoplasma when obtaining cells from an outside source.
• Test for mycoplasma every 3 months.
• Test every month if sharing hood space or a CO₂ incubator with another person.
• Be vigilant if experiment results are not consistent/reproducible.

The Challenge of Mycoplasma Detection

Mycoplasma has long been recognized as a common contaminant of cell cultures, but infected cells may go undetected for years. Among the available detection methods (broth culture, fluorescence staining, bioluminescence, ELISA, and PCR), PCR is the most sensitive, specific, and convenient method⁴. It is used by most mycoplasma detection kits, such as those from Thermo-Fisher, Systembio, Sigma, and ATCC. However, all of these PCR kits require DNA extraction or sample preparation. Post-PCR gel electrophoresis is also required, except for real-time PCR kits. Therefore, current kits have, until now, involved a long and laborious process.

A simple, sensitive, and quick tool is needed.