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mycoplasma and fungus


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#1 kant0008

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Posted 07 July 2004 - 05:11 PM

Hi all!
I'm trying to find out about fungul and mycoplasma contamination. The other day I gave some of my cells to a collegue who is studying fungus, as a negative control, yet my cells came up positive in his pcr. he seems quite certain that it's a not a false positive. My cells seem fine, and I can see no usual fungal "characteristics"- long filaments or lawns. However there are a few floating "objects" in the flasks- they are not of a regular shape, can form clumps, about a quarter of the size of my cells (so too big for bacteria). I can't figure out if they multiply at all, there seem to be a few at all times, with a small increase in number over time- but what I am not sure about is if they multiply or if some of my cells are dying and that's just debris. Can it be mycoplasma? Woudl I be able to see it under a microscope? Is there any way to treat this infection?
At the moment I'm adding 4 different antibiotics to these cultures, as they are transfectants and I'm keeping two plasmids, so the usual Pen/strep mixture, plus neomycin and hygromycin (that's why I was surprised to get an infection, if that's what it is, and also think that maybe my cells dye faster with all tese antibiotics).
Any comments please?

#2 mujan

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Posted 09 July 2004 - 11:05 AM

check out BM cyclin. can also try DAPI to stain extranuclear DNA. sometimes it is hard to see. you need a good microscope and maybe 63X objective.

#3 phdconsult

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Posted 11 July 2004 - 06:04 PM

You might be dealing with a Nocardia Asteroides or Mycobacterium gordonae contamination. Try supplementing with 5ug/ml of p-aminosalicylic acid or Isonicotinicacid hyrazide or ethambutol or D-cycloserine. These drugs should kill of the contaminant, yet not affect other species.




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