Mycoplasma Detection Service

With the increased use of cell cultures contaminations are increasingly frequent, i.e. cellular contamination (cross-contamination between cell cultures) and contamination with microorganisms (microbial contamination). While bacterial and fungal infections of cultures are relatively easy to detect, to prevent and to treat, contamination with Mycoplasma represents a much bigger problem in terms of incidence, detectability, prevention, eradication and effects. It was estimated that between 5 and 35% of cell cultures in current use are infected with Mycoplasma.

• Why Test for Mycoplasma Contamination?
• PCR-based Mycoplasma Testing
• Applications
• How To Prepare Your Samples
• What type of report will be given to you?
• Contact Person

Why Test for Mycoplasma Contamination?

Mycoplasma contamination of cell cultures can:

• Affect cell growth and morphology
• Inhibit cell metabolism
• Disrupt DNA and RNA synthesis
• Produce chromosomal aberrations
• Change antigenecity of cell membranes
• Mimic virus infection
• Alter virus yields

PCR-based Mycoplasma Testing

PCR is the fastest and most sensitive method of Mycoplasma detection available. We offer a nested (two stage) PCR-based Mycoplasma test that includes a negative control and a positive control in each test.

Applications

With our PCR-based assay, we can detect as few as one colony-forming unit (CFU) per sample. It detects >120 different Mycoplasma species, including the eight most commonly encountered Mycoplasma contaminants in cell culture.

How To Prepare Your Samples

It is recommended to perform 2 passages in media free of antibiotics and other additives before sample submission. Cell cultures should be confluent at the time of harvest. It is recommended that it be at least 3-4 days since the last feed/medium change. Prepare sample(s) following one of the procedures bellow

•  For adherent cells:  The cells should be harvested by scraping into conditioned media. Trypsin should not be used. (Culture medium without cells can also be tested; however, this is not recommended(
• For cells in Suspension: Count cells. Medium should contain cells at ~10⁴ - 10⁵ /ml.

For each tested sample prepare 2 replicates of 1 ml each (1 for testing and 1 for retain to be used for retest if needed according to the test acceptance/rejection criteria). Aseptically place 1 ml sample in polypropylene screw-capped vial(s), allowing some air space for expansion (cells are frozen in conditioned medium without cryopreservative)

If you are submitting more than one cell culture sample, put samples for each culture in a separate bag.

What type of report will be given to you?

We will provide you with a result form – stating whether your cell culture samples are positive or negative for the presence of Mycoplasma.