Mycoplasma species are bacteria that readily contaminate cell cultures leading to serious reproducibility and quality issues in cell biology research and bioprocessing. Their small size, lack of a cell wall, and resistance to commonly used antibiotics makes Mycoplasma contamination notoriously difficult to detect and persistent. It is estimated that 15-35% of cell lines worldwide are contaminated with Mycoplasmas. Flip Mycoplasma represents a powerful new tool for the rapid detection of Mycoplasma contamination in cell cultures.

Flip Mycoplasma has been demonstrated experimentally to detect the six Mycoplasma species responsible for the vast majority (>95%) of incidences of cell culture contamination: M. arginini, M. orale, M. fermentans, M. hominis, M. hyorhinis and A. laidlawii. in silico analysis has further demonstrated inclusivity of the test for >100 other Mycoplasma species which are less frequent contaminants of cell culture.

No. Flip Mycoplasma is for research use only. It is not for use in diagnostic procedures.

Flip Mycoplasma is a single-use (disposable) nucleic acid amplification test for the qualitative detection of 16S ribosomal RNA from the mycoplasma species that contaminate cell culture samples. The test is provided in a ready-to-use format containing lyophilised reagents for the amplification assay and a detection strip to report the test results.

Flip Mycoplasma is provided in a convenient lyophilised format for stable long-term ambient storage. Store Flip Mycoplasma including all test components at ambient temperature. Do not freeze. In case of refrigeration or other exposure to cold temperatures, ensure that the test is allowed to come to ambient (room) temperature prior to use.

Flip Mycoplasma can be used to test cell culture supernatant from suspension or adherent cell cultures prior to passaging or changing media. It can also be used to screen fetal bovine serum (FBS) for Mycoplasma contamination prior to its use in the supplementation of cell culture media. For adherent cell cultures, obtain supernatant directly from the cell culture flask. For suspension or semi-adherent cultures, first remove cultured cells from the supernatant by centrifugation at 500g for 5 min.

Flip Mycoplasma has been found to function effectively in a wide range of cell culture supernatant samples prepared with different media, cell types and additives. Whilst it is not possible to evaluate the test performance with every type of sample given the wide diversity of cell culture work, Flip Mycoplasma has been found to have excellent robustness to components that may be present in cell culture supernatant samples. The test is intended for use prior to passaging or changing media. Cell cultures that are grown to a high degree of confluence may accumulate inhibitors over time that negatively impact the performance of the test.

Laboratory pipettes and tips

Preheated heat block or thermal cycler suitable for 0.2ml tubes, set to 50°C

1.5ml microcentrifuge tubes

Timer

Microcentrifuge (for suspension or semi-adherent cell cultures only)

Samples must be diluted at 10% (v/v) in the Sample Buffer provided with the test according to the test instructions or quick use guide. Other buffers are not compatible with Flip Mycoplasma and it is not possible to test samples directly.

To ensure optimal test performance, do not remove the foil from the Assay Tube until just before use. After adding the prepared sample to the Assay Tube proceed immediately to perform the remaining test steps according to the instructions or quick use guide.

The Flip Mycoplasma test can be heated using any laboratory heat block or thermal cycler suitable for 0.2ml tubes, or the USB-C powered Flip Nest (contact support@fuse-dx.com for further information). We do not recommend using a water bath, which can be a source of contamination.

The heat block or PCR machine should be preheated to 50°C prior to performing the test. Use of an incorrect temperature may impact the performance of the test. If you believe that an incorrect temperature has been used then it is recommended that the test is repeated.

To ensure optimal performance the assembled Flip device should be incubated for exactly 10 minutes using a timer. After 10 minutes, remove the test from the heat and flip the test according to the instructions or quick use guide. Incubating the test for less than or greater than 10 minutes may impact the performance of the test. If you believe that an incorrect incubation time has been used then it is recommended that the test is repeated.

If no liquid is seen flowing on the test strip 30 seconds after you flip and gently shake the cassette downwards, then repeat this step.

The test results can be read from 2 minutes after the fluid is observed flowing onto the test strip by observing lines on the test strip through the readout window. The test results should not be read early but can be read any time after 2 minutes.

Flip Mycoplasma contains an internal process control that has been designed to control for assay reagent function and sample inhibition. The appearance of the control line on the test strip indicates that the assay reagents maintained their functional integrity and the sample did not significantly inhibit assay performance.

The absence of a line in the control position (top) indicates an invalid test. An invalid test may have a number of potential causes. It is recommended that an invalid test is repeated. If the repeat test is invalid then contact support@fuse-dx.com for further assistance.

The presence of any line in the results position, even a very faint line, indicates a positive test result. The test is a qualitative (yes/no) test and does not provide the quantitative value of detected organism present.

External controls are not provided with Flip Mycoplasma. We recommend the use of M. orale 10CFU™ Sensitivity Standard (Cat. No. 102-2003, Minerva Biolabs GmbH). See the test instructions for information on how to perform external positive and negative controls.

Prepared samples are stable for at least one hour at room temperature, 24 hours at refrigerated temperature (2-8°C) or one month frozen (-20°C). Ensure that the prepared sample is allowed to warm to room temperature prior to use.

Yes prepared samples can be retested, provided they have been stored under the recommended conditions.

The Assay Tube and Readout Cassette are single-use. Do not attempt to reuse the Assay Tube or Readout Cassette. After use, the test contains large amounts of amplified target (amplicon). Do not disassemble the Assay Tube and Readout Cassette, as this is highly likely to lead to amplicon leakage and contamination of the use environment which could impact the results of further tests.

If a cell culture is found to be positive for Mycoplasma it is recommended that where possible the contaminated cell culture should be discarded and grown again from a fresh, uncontaminated stock. However, if the cell line cannot be easily replaced, a variety of protocols and reagents exist for the removal of Mycoplasma from a live cell culture. Consult your local distributor for products that they offer or contact support@fuse-dx.com.  

If you suspect that the test environment has become contaminated then it is recommended to perform a negative external control test as described in the test instructions. The test environment and any equipment used for testing can be cleaned with a suitable laboratory detergent before repeating a negative external control test. 

Used Flip Mycoplasma tests should be discarded with other laboratory waste, according to local procedures. The outer cardboard packaging, instructions and quick use guide can be recycled. The Assay Tube and Readout Cassette are single use. Do not disassemble the Assay Tube and Readout Cassette, as this is highly likely to lead to amplicon leakage and contamination of the use environment which could impact the results of further tests. Can I store prepared samples for the Fuse Mycoplasma test?

Flip Mycoplasma is not classified as hazardous. The Safety Data Sheet for this Product is available on www.flip-dx.com/mycoplasma or contact Fuse Diagnostics Ltd (support@fuse-dx.com). The Sample Buffer may contain irritants. Do not ingest the contents of the tube. If the contents of the tube are splashed in your eyes, flush your eyes with water. If the contents splash onto your skin, wash with soap and water. If irritation persists, consult a healthcare provider.

Please see www.flip-dx.com/mycoplasma for more information. Contact your local distributor or Fuse Diagnostics Ltd (support@fuse-dx.com) with any questions or comments.