Re: Mycoplasma Detection

From: Simon_Q_Rice@sbphrd.com
Date: Thu Feb 25 1999 - 03:55:32 EST


Elizabeth,
Assuming the PCR conditions were controlled and you know reagents, method
of preparation of the sample etc are ok, I'd be surprised if this technique
wasn't the most sensitive of the two you've described-it seems strange then
that the least sensitive method should give you a positive signal.

My suggestion would be to check conditions of the PCR with a positive
control, that cell cultures have been grown in antibiotic free medium for
at least three passages (that includes selection agents such as
neomycin/geneticin which have anti-microbial properties) and then recheck
alongside another technique.  We have used the Genprobe detection kit for a
number of years and it works very successfully-however the PCR should
remain the most sensitive.

I hope this is useful.

Regards

Simon

Simon Q.J. Rice
Gene Expression Sciences
Smithkline Beecham Pharmaceuticals,
NFSP-N, Harlow, UK.






et.kraus@baylordallas.edu on 22-Feb-1999 22:55



To: cyto-inbox
cc:    (bcc: Simon Q Rice)
Subject:  Mycoplasma Detection





This message is being posted for a co-worker.  Not necessarily flow-related
but could be useful to the group:


Does anyone have experience in mycoplasma detection in cell lines?

Our dilemma:  PCR results (ATCC Mycoplasma Detection Kit) are negative
while
confocal microscopy detects positive DAPI labeling in the cytoplasm of the
same cell line.

Any help would be appreciated.

Elizabeth T. Kraus
et.kraus@baylordallas.edu



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