I'm curious whether anyone on here has experience with InvivoGen's PlasmoTest to detect Mycoplasma infection of cell culture lines, as well as perhaps how it compares with PCR and Hoechst staining.
I know the gold standard is direct culture, but at best we would just outsource that to a commercial service.
I havn't used that particular kit, which quite frankly seems like a bit of a pain to use on a regular basis -- although it's nice that it is colorimetric.
For our routine screening of culture media we used a Lonza kit, MycoAlert:
http://www.lonza.com/products-services/bio-research/cell-culture-products/mycoplasma-detection-and-removal/mycoalert-mycoplasma-detection-kit.aspx
This one was very easy to implement, so there was no excuse to not have a regular screening program. It's an enzyme based luminescence assay. Lonza recommends 100uL of media and get results in about 30min, you just need access to a luminometer (or of course they can sell you one).
Lonza claims that the sensitivity is 50 CFU/mL, similar to direct culture or PCR, which seems a bit hard to believe -- that's about 10X better than the PlasmoTest.
Depending on your luminometer, you can cut the sample and reagent volumes and thus get more assays out of the kit.
I use Lonza kit, MycoAlert too. IT works well, but you need a luminometer
Hi Kameron,
I found that Bioreliance has a great webpage about the different methods/approaches to look for mycoplasma. http://www.bioreliance.com/mycoplasmadetection.aspx I've attached a PDF file from their site that goes into great detail comparing methods.
I personally have not used the InvivoGen kit but have heard good things.
iQ Biosciences offers the PCR based method which seems to work fine for a quick screen. http://www.iqbiosciences.com/bioservices/testing-services/mycoplasma-testing.
Best Regards,
Omar
We used Lonza's MycoAlert kit today to screen cell lines routinely used in our lab. Results came out on expected lines. As suspected HEK 293 and BHK-21 were mycoplasma contaminated; insect cell lines, FBS (Biological Industries, Israel), adult bovine srum (Sigma Aldrich) were mycoplasma free. Only drawback is it requires Luminometer. On prior appointment Lonza provides Luminometer service at our lab free of cost.
If you want to outsource, then I would send the cells to Multiplexion. They can do the Authentication and contamination testing (Mycoplasma, interspecies and viral contaminations) at very affordable prices.
http://www.multiplexion.de/en/
ResearchGate notified me that this got 150 reads, so I figured I'd post an update in case it's helpful for the future.
We decided to continue using PCR for detection, with some modifications for ease of use. For your convenience I'm attaching our protocol here. It largely references a Nature Protocols paper, but we made some modifications to speed it up, notably using an all-in-one mastermix and loading buffer that really makes it easy to run lots of samples at once.
We've been using for years the PlasmoTest by Invivogen. For us it works nice and you can show the results to the users of the unit of cell culture because with the MW96 that you use to perform the test, people can see the difference between the negative control and their cells if they have any contamination.
Kind regards.
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