Have any of you ever used the mycoplasma-detection kit by Invivogen based on the HEK-Blue-2 cell line and reagents? Can I compare its accuracy with a mycoplasma-PCR? If anyone has experience with this kit, please share.
The aim is to have a mycoplasma detection method which is not based on PCR, we do not have a cycler or enough room for all the gel-casting and so on.
Dear Alexander,
Last year, we've had some problems of unexplained cell death in our cell cultures. I have suspected that our cell lines are infected with mycoplasma because these problems occured in several labs at the same time. The kit that was used for mycoplasma detection was from Invitrogen: MycoFluor™ Mycoplasma Detection Kit, Ref. M7006. The mycoplasma are then observed by fluorescence microscopy as small blue dots, which probably correspond to the mycoplasma DNA (it is not explained by the kit manual). I do not think that its accuracy is comparable to that of mycoplasma-PCR, but it allowed us to detect their presence in our cultures.
The cells were then treated with a compound from Invivogen (Plasmocin™ treatment, Ref. ant-mpt), and nowadays we can say that our cell cultures are clean. However, it took several months to obtain this result.
We have also replaced one of the cell lines (it was bought), which seemed not sensitive to the treatment. Unfortunately, it is not possible to replace all of them because it is too expensive.
The lab was also disinfected with a specific compound (Mycoplasma-Off - Surface Disinfection Spray, Ref. 15-1000) bought from Lucron ELITechGroup.
I hope this may help you, but please do not hesitate to ask me further information.
Kind regards,
Carmen
Beacause of the suppression of Mycotect kit from GIBCO? we have had to select a new kit. We tested all sots of principles available and have been chosen this on because of its efficiency in detecting mycoplasmic contamination (is is more sensitive than PCR based tests). We have "miniaturised" the test and it is really easy to use in a cell culture laboratory.
We have used the Invivogen PlasmoTest and it is very good and straightforward to use and due to its 96 well format space is not a constrain (however you will need the incubator space to grow the HEK-Blue-2). We have not compared it directly with a PCR based test but other colleagues had and preferred it. The key issue with this test is to make sure the HEK are in selection for a least a few days prior to the test and they are healthy, otherwise the media colour change might not be strong enough (however the positive controls supplied will indicate if this is the case). Please let me know if you have any other questions.
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