The presence of mycoplasma in cell culture is highly undesirable and can conclude in a serious distortion or even failure of in vitro experimental results coming from:
The method involves isolating DNA from a sample of a conditioned eukaryotic cell medium. Subsequently, amplification and detection of a highly conserved region encoding 16S ribosomal RNA specific for mycoplasmas is performed by real-time PCR. This method is used for the qualitative determination of mycoplasmas in a sample.
Test parameters
The method used meets the requirements of the European Pharmacopoeia given in Chapter 2.6.7.
Limit of detection: 10 cfu/ml
Requirements for the material to be tested
The customer provides us with 0.5 ml of cultivation medium (or cells suspension), prepared according to the instructions below.
Preparation of cells to be tested
To ensure reliable test results, please keep carefully to the recommendations as follows:
- Cell confluency must be at least 80 % at the moment of medium withdrawal.
- Withdrawal of a medium sample to be tested must be performed no sooner than 2 days following the last exchange of medium.
- Incubate at least 0.5 ml of the collected culture medium at 95 ° C / 10 minutes. Then centrifuge (11,000 × g, 15 seconds) and store at -20 ° C.
- In the case of suspension cell testing, a maximum concentration of 1×10^6 cells / ml is required.
Sample delivery
If the delivery time does not exceed 24 hours, send samples chilled to 2 to 8 °C (on wet ice), otherwise we recommend delivery on dry ice.
Output of the test
We provide you with a protocol in English with a statement if the presence of mycoplasma was detected.
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