summary of mycoplama detect

Richard Near rocket1 at bu.edu
Thu Jul 28 21:51:17 EST 1994


Here is a crude set of extracts from the letters I received concerning
kits available to detect mycoplama contamination:


Check out Ian Freshney's Animal Tissue Culture Techniques (any of the 3
editions)--it will have a staining procedure that's much cheaper than PCR. Or
look at a package insert from Boeringer Mannheim's BM Cyclin (anti-mycoplasma
antibiotic regimen--works great for me!)--it has a DAPI staiing protocol that
takes about 5-10 minutes.  When my cultures were contaminated there was *no*
doubt when I compatred (make that *compared*) an uncontaminated culture.
]Hope this helps,
Beverly
Date: Thu, 14 Jul 94 10:27:58 EDT
From: plc at med.unc.edu (Philip L. Carl)
To: rocket1 at bu.edu
Subject: Re: Mycoplasma detection kits?
Newsgroups: bionet.molbio.methds-reagnts
Organization: UNC-CH School of Medicine
Status: O

Strategene sells a PCR based kit.  Haven't tried it.

Phil Carl

In my experience, the best detection that I have found for mycoplasma is by 
using a kit from a company called Gen-Probe.  They are in San Diego phone 
800-523-5001.

The principle is hybridization to mycoplasma rRNA.  All very step-by-step and 
sensitive.

Bob Jackman
Richard,
        we are using the mycotect kit available from Gibco BRL
Life Technologies. The kit utilises the fact that mycoplasmas 
contain large amounts of adenosine phosphorylase which converts
a non toxic adenosine analogue(6MPDR)to a toxic one. All you have to
do is culture up some 3T6 cells add supernatant from your murine
cultures, plus the 6MPDR and wait a couple of days. If your cells
are mycoplasma infected the 3T6 cells will be totally wiped out.

They reckon that this method detects all major species of mycoplasma
and we use it routinely to monitor our Mab secreting cell lines.

I tried the Hoechst stain but I couldnt tell the difference between
the positive and negative controls!

I have heard of the PCR test but nobody I know uses it.

I HOPE THIS IS OF USE
PLEASE EMAIL ME IF YOU CAN THINK OF OTHER SUGGESTOINS

RICK
ROCKET1 at BU.EDU





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