ECACC offers a range of tests for the detection of microbial contamination within cell cultures and related culture media and reagents
Both pharmacopoeia and in-house detection assays are available to detect mycoplasma, bacterial, fungal and viral contaminants.
Tests are performed in its dedicated quality control laboratories away from its routine cell culture and banking facilities.
Testing for mycoplasma should concern everyone working with cell cultures.
A mycoplasma infection of a cell culture can seriously affect the performance of the cell line and its associated research results.
The presence of mycoplasma can cause alterations in the rate of cell growth, induction of morphological changes, chromosomal aberrations, changes in amino acid and nucleic acid metabolism, induction of transformation, induction of apoptosis, induction of cytokines and oxidative radicals, macrophage activation, inhibition of antigen presentation, and induction of signal transduction.
Therefore, the research results obtained using a cell line contaminated with mycoplasma can vary widely from those generated using a mycoplasma free cell line.
In addition to the effects mycoplasma can have on an individual cell line, the introduction of a contaminated culture can devastate a cell culture facility due to its ability to spread rapidly.
Therefore, it is vital that cell cultures are tested both at the start of and at frequent intervals during their propagation.
Three mycoplasma detection methods are routinely offered by ECACC as reliance on a single detection method for anything other than screening purposes is not advisable.
If a cell line has not been tested for some time, ECACC recommends that all three tests are undertaken to determine the mycoplasma status of the cell line.
Cell lines and culture media can be tested for mycoplasma contamination using one, or a combination of techniques.
These methods are routinely used at ECACC for the detection of mycoplasma.
Culture and DNA stain are the only two tests recognised by the FDA.
As not all mycoplasma grow in culture and DNA stain only has a sensitivity of 104cfu/ml, it is recommended that both methods are used to determine whether a sample is free of mycoplasma.
ECACC also tests for mycoplasma using PCR analysis.
Currently this technique is not recognised by the regulatory bodies.
PCR can give a quick indication of a mycoplasma positive cell line.
However, a PCR negative result does not guarantee that a test sample is mycoplasma free and it is recommended that culture and DNA stain tests are carried out in conjunction with PCR to confirm the result.
