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Mycoplasma contamination is subtle and pervasive

Mycoplasma are small (0.15 µm-0.3 µm) bacteria with no cell wall and a flexible membrane. These characteristics of Mycoplasma mean that contamination is subtle. Their lack of a cell wall and flexible membranes make them difficult to detect using microscopes, and their small size allows them to proliferate to high concentration within mammalian cell culture without any noticeable turbidity or other change in cell culture appearance.
Estimates suggest that between 5% and 30% of the world’s cell lines may be contaminated with one or more of the several known species of Mycoplasma.

Mycoplasma contamination is difficult to avoid because the major sources of contamination are lab personnel, standard cell culture components (for instance, fetal and newborn bovine serum and swine-derived trypsin), and cross-contamination from other cell cultures.

Contamination of your culture may not be apparent until your cell line is lost or your antibody is tested.


What is happening in your cell culture that you can’t see?


Mycoplasma contamination affects cell culture systems in many ways. Some effects may include:

  • Change of gene expression/protein expression profiles
  • Inhibition of protein biosynthesis due to arginine starvation
  • Inhibition of cell proliferation due to starvation/competition for nutrients in media
  • Production of toxic metabolites or nutrient competition adversely affecting antibody production
  • Changes in cell membrane antigenicity
  • Induction of apoptosis
  • Penetration of host cells, leading to long-term contamination
  • Acidification of media due to glucose metabolism
  • Chromosomal abnormalities
  • Inhibition of cellular differentiation
  • Activation of immune cells
  • Inhibition of cell growth
  • DNA fragmentation due to nucleases produced by mycoplasma
  • Compromised production of viruses
  • Inhibition of cell metabolism
  • Reduction of transfection efficiency


The best treatment is prevention

Cross-contamination from other contaminated cell lines is one of the main sources of mycoplasma contamination. Elimination of mycoplasma contamination from a culture is almost impossible; mycoplasma is resistant to most antibiotics and can survive liquid nitrogen without cryopreservation techniques, contaminating other cells stored in liquid nitrogen.

The best way to prevent whole-scale mycoplasma contamination of your antibody-producing clones:

  • Routinely test your existing cell lines and discard any positive cultures
  • Routinely quarantine and test any new cultures from outside sources


Microsart® Mycoplasma Detection Kits, regulation-compliant (European Pharmacopeia 2.6.7)

Other qPCR-based Mycoplasma detection kits on the market don’t give you an answer in three hours that

  • is as efficient- minimize pipetting effort by using less tubes (test 10 samples with 12 tubes using Microsart Mycoplasma Detection Kits vs 24-28 tubes for other kits. Directly incorporate controls in to the sample mix
  • is as cost-effective
  • uses your existing real-time PCR intstrument

Microsart® Mycoplasma Detection Kits offer qPCR-based detection with an answer in three hours using

  • highly-specific TaqMan® probes to reduce the risk of false-positives
  • multiplex PCR for sample and inhibition control to reduce the risk of false-positives and save on handling and analysis
  • inactivated, noninfectious validation standards to reduce the risk of cross-contamination

Download Kit Flyer


Sartorius also offers non infectius validation standards for several mycoplasma species.

  • Mycoplasma arginini (quantified CFU standard)
  • Mycoplasma orale (quantified CFU standard)
  • Mycoplasma gallisepticum (quantified CFU standard)
  • Mycoplasma pneumoniae (quantified CFU standard)
  • Mycoplasma synoviae (quantified CFU standard)
  • Mycoplasma fermentans (quantified CFU standard)
  • Mycoplasma hyorhinis (quantified CFU standard)
  • Acholeplasma laidlawii (quantified CFU standard)
  • Spiroplasma citri (quantified CFU standard)
  • Mycoplasma arginini (quantified DNA standard)
  • Mycoplasma orale (quantified DNA standard)
  • Mycoplasma gallisepticum (quantified DNA standard)
  • Mycoplasma pneumoniae (quantified DNA standard)
  • Mycoplasma synoviae (quantified DNA standard)
  • Mycoplasma fermentans (quantified DNA standard)
  • Mycoplasma hyorhinis (quantified DNA standard)
  • Acholeplasma laidlawii (quantified DNA standard)
  • Spiroplasma citri (quantified DNA standard)

Mycoplasma cell

Microsart® Mycoplasma Detection Kits: a kit to meet your antibody-discovery program’s needs

Microsart® RESEARCH Mycoplasma Detection Kit

Fast and reliable detection of Mycoplasma DNA in cell culture supernatants for research and development labs

Download RESEARCH Datasheet

Microsart® AMP Mycoplasma Detection Kit

Reliable and sensitive detection of DNA from more than 70 Mycoplasma species, validated for sensitivity, specificity, and robustness according to EP 2.6.7 for in-process control and lot release

Download AMP Datasheet

Want to learn more or request a demo?

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