Validation of MycoGenie Rapid Mycoplasma Detection Kit

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Mycoplasma | General Overview

Mycoplasma contamination is a significant issue for cell culture scientists and can have a severe impact on protein production & cell health. They can be transmitted by aerosols from cultured cell lines, contaminated tissue, cultured media and biological waste such as carcasses, aborted fetal tissue and embryos that have been infected or not treated with antibiotics.

Mycoplasma contaminated cell cultures tend to be discarded which can result in a severe economic loss for the bio-pharmaceutical industry. Alternatively, scientists can use newly developed elimination reagents such as the Assay Genie MycoGenie MycoPlasma Elimination Kit that removes Mycoplasma without the need to discard precious cell lines and cultures.

Cell Culture Contamination

Mycoplasma-infected cells produce sub-optimal amounts of protein and increase the rate at which those cells die. Therefore, Mycoplasma contamination can reduce both the quantity and quality of protein produced by cultured cells.

Mycoplasma contamination can rapidly reach very high levels & infected cell culture monolayers often resemble tumour growth.

These infected cell culture monolayers are often responsible for aerosolization of Mycoplasma and subsequent contamination of cell culture equipment, plasticware and cell lines.

Assay Overview

Assay Genie MycoGenie Rapid Mycoplasma Detection Kit detects Mycoplasma in cell culture using a visual determination method. The kit utilises a highly sensitive isothermal amplification method coupled with a colorimetric readout that can detect up to 28 Mycoplasma species, including 8 of the most common species associated with cell culture contamination.

Assay Features

• Rapid | Detect Mycoplasma in 1 hour (28 species detected) with Isothermal amplification & pH color change that’s easily read by eye.
• Simple | Easy protocol with detection by visual determination of color change meaning no expensive equipment needed.
• Sensitive | Detect as little as 500cfu Mycoplasma per 1ul of cell culture supernatant.
• Flexible | Detect Mycoplasma in adherent & suspensions cells such as Vero, MDCK, SP2/0, 293T, HepG2. HeLa, A549, MB-MDA231, L929, MEF, CHO, NS0, 293F, mouse hybridomas, Sf9, BHK21 & more.
• Compatible | With a wide selection of cell culture media & sera such as Fetal bovine/calf serum, horse serum, Gibco KSR serum replacement & CD FortiCHO, CDM4, Expi 293 Medium, CD Hybridoma, Grace, DMEM, 1640, F12 & more.

Conclusion

In conclusion, the sensitivity of the experimental protocol for qPCR is equivalent to that of isothermal amplification driven by the the MycoGenie Rapid Mycoplasma Detection kit. In detecting Mycoplasma both were able to detect Mycoplasma in stock solutions with a copy number in the hundreds. The PCR amplification protocol has lower sensitivity and was only able to detect Mycoplasma in stock solutions with a copy number in the thousands:

• The limit of detection for PCR was 3.07 x 10^3 copies of Mycoplasma.
• The limit of detection for qPCR was 3.07 x 10^2 copies of Mycoplasma.
• The limit of detection for the MycoGenie Rapid Mycoplasma Detection kit isothermal amplification was 3.07 x 10^2 copies of Mycoplasma.

For more information on the mycoplasma detection kit, please contact one of our account managers.