Why Test for Mycoplasma? 

The term “mycoplasma” refers to a class of bacteria that is commonly present in cell culture^(1-2). More correctly known as mollicutes, these bacteria lack a cell wall, making them resistant to commonly used antibiotics such as penicillin⁽³⁾. Mycoplasma are easily spread through droplets during cell culture and can survive in dried form for months^(4-5).

Mycoplasma contamination is difficult to see with a microscope and requires specific testing to be detected⁽³⁾. Contamination can have a profound impact on experimental work. Depending on the species present, mycoplasma organisms can affect cell growth, metabolism, tumourigenicity, and response to chemotherapy^{(1, 6-7)}.

Mycoplasma Testing at CellBank Australia

Mycoplasma testing is performed on samples of culture medium. We recommend that cells are grown in antibiotic-free medium for at least two weeks and/or passaged at least twice. Media should not be changed for 3-4 days prior to sampling. Antibiotics can suppress bacterial contamination, which means that growing cells in antibiotics can increase the risk of false negative results.

A sensitive in-house PCR method is used to detect mycoplasma DNA, with an internal control to detect PCR inhibition, which can affect culture samples.

The MycoAlert® Mycoplasma Detection Kit (Lonza) is used as a second method to detect viable mycoplasma, based on enzymatic activity.

A third test method can be used if results are unclear. Enrichment can be performed using an indicator cell line, resulting in rapid growth of low levels of mycoplasma that might be difficult to detect in the original test sample.

Price:    From $150 per sample, excluding GST.

Terms and Conditions

Please note that all CellBank Australia services are suitable for research purposes only and are not appropriate for human clinical or diagnostic samples. CellBank Australia can only test cell lines classified as Physical Containment Level 1 (PC1) and Level 2 (PC2). Full terms and conditions can be found in the ‘Attachments’ tab.

Please be reminded that we cannot guarantee detection of all species of Mycoplasma. Low levels of Mycoplasma may arise through chronic infection masked by antibiotic use. If antibiotics are used, testing should be performed after 2-4 weeks’ growth in antibiotic-free medium.

Orders:

At present, we do not accept orders online for this service. To request a Mycoplasma testing:

• Contact us using the ‘Ask a Question‘ tab and tell us about any special needs that you may have.

• Download a copy of the ‘Request for Mycoplasma Analysis’ form, available from the ‘Attachments’ tab.

• Complete and return the form via email (cellbank@cmri.org.au).

• Upon confirmation, prepare your samples and ship them to CellBank Australia in accordance with the instructions on the Request form.

References

1. Rottem S, Barile MF (1993) Beware of mycoplasmas. Trends Biotechnol 11(4): 143-51. PMID: 7763647.

2. Razin S, Hayflick L (2010) Highlights of mycoplasma research-An historical perspective. Biologicals 38(2): 183-90. PMID: 20149687.

3. Drexler HG, Uphoff CC (2002) Mycoplasma contamination of cell cultures: Incidence, sources, effects, detection, elimination, prevention. Cytotechnology 39(2):75-90. PMID: 19003295.

4. McGarrity GJ (1976) Spread and control of mycoplasmal infection of cell cultures. In Vitro 12(9):643-8. PMID: 1034618.

5. Nagatomo H, Takegahara Y, Sonoda T, Yamaguchi A, Uemura R, Hagiwara S, Sueyoshi M (2001) Comparative studies of the persistence of animal mycoplasmas under different environmental conditions. Vet Microbiol 82(3):223-32. PMID: 11470544.

6. Namiki K, Goodison S, Porvasnik S, Allan RW, Iczkowski KA, Urbanek C, Reyes L, Sakamoto N, Rosser CJ (2009) Persistent exposure to Mycoplasma induces malignant transformation of human prostate cells. PLoS One 4(9):e6872. PMID: 19721714.

7. Vande Voorde J, Sabuncuoglu S, Noppen S, Hofer A, Ranjbarian F, Fieuws S, Balzarini J, Liekens S (2014) Nucleoside-Catabolizing Enzymes in Mycoplasma-Infected Tumor Cell Cultures Compromise the Cytostatic Activity of the Anticancer Drug Gemcitabine. J Biol Chem 289(19):13054-65. PMID: 24668817.