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Kit for Mycoplasma detection by PCR


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9 replies to this topic

#1 Tim

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Posted 24 July 2002 - 01:56 AM

Can anyone recommend the best commercially available kit for Mycoplasma detection by PCR.

#2 Filbert

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Posted 24 July 2002 - 05:31 PM

Try to visit A.T.C.C homepage: http://www.atcc.org

#3 mujan

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Posted 26 July 2002 - 09:32 AM

There are companies that will do the diagnostic for you.  It may be better since they have all the controls and assays set up.  The NIH uses this system before any new cell lines are permitted into certain labs.  There are many strains of mycoplasm that your PCR approach may not pick up.  Gibco/invtrogen has a cell based kit and (maybe) a PCR kit.  There is a company in upstate New York that will clean your cells up for a fee. I can't remember their name.  They have several set ups ranging from the high hundreds to a about 1500.  The expensive system uses a mammalian host to harbour your cells. The host immune system will kill off the mycoplasm.  Check the Web using the go.com search engine.  Type in mycoplasm.  Alternatively, try BM-Cyclin from boehringer mannheim.  The protocol is cheap, easy, and very effective.  You'll have clean cells in about 3-4 weeks.

#4 Paola PC

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Posted 01 August 2002 - 08:54 AM

After frustrating staining of coverslide grown cells(DAPI and Hoechst33258), I am now using the PCR ELISA mycoplasma detection kit from Roche with good results! Good luck

#5 marcbaeuerle

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Posted 08 March 2005 - 03:02 AM

Minerva Biolabs provide superior PCR detection kits for mycoplasma.
Very simple, reliable and inexpensive.
www.minerva-biolabs.com :D

#6 desertrose

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Posted 07 April 2010 - 09:01 AM

Are these kits suitable for detection of mycoplasma detection in tissue cultures used for vaccine production

#7 bob1

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Posted 07 April 2010 - 05:07 PM

PCR is not sensitive enough and provides too many false readings (positive and negative) for it to work reliably - you need to use the Hoescht staining kits and soft agar assay for decent results!!!

#8 rhombus

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Posted 08 April 2010 - 03:50 AM

PCR is not sensitive enough and provides too many false readings (positive and negative) for it to work reliably - you need to use the Hoescht staining kits and soft agar assay for decent results!!!




Oh my God not again !!!!!!!!!

bob1 is completely right as usual.....READ THE ATCC WEBSITE......I have copied the relevant bit....please see below:-


Testing cell cultures for mycoplasma is essential to ensure reliable research and biotechnology products. Mycoplasma contamination does not produce turbidity but can have adverse effects on cell line growth and characteristics.

With ATCC's Mycoplasma Testing Service, DNA staining and direct culture methods are performed simultaneously for each sample. Together these methods can detect mycoplasma infection for most of the known species. Each test includes positive and negative controls and complete testing requires four to five weeks. ATCC no longer offers PCR-based mycoplasma testing.


For the un-initiated " ATCC NO LONGER OFFERS PCR-BASED MYCOPLASMA TESTING"

The FDA only approve 2 tests: Hoescht Staining in combination with soft agar growth assay.

The reason for dropping PCR is that as bob1 rightly states, you get too many false negatives.....and this is the main problem....FALSE NEGATIVES.


The ATCC are the worlds largest supplier of cell lines and hybridoma's..... I think they know what they are talking about.


Uncle Rhombus.

#9 desertrose

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Posted 08 April 2010 - 07:56 AM

great answers thanks a lot

#10 rhombus

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Posted 09 April 2010 - 03:01 AM

Are these kits suitable for detection of mycoplasma detection in tissue cultures used for vaccine production



Sorry but missed your post completely. My first job ever was to produce Human Vaccine in cell culture (Polio Vaccine). The test used then for Mycoplasma Detection was the soft agar growth assay......and 32 years later it is still the same. As I quoted before, the Food and Drug Administration (FDA), has only approved Agar and Hoescht staining for Mycoplasma detection. If it is good enough for vaccines and drugs then that is the ultimate test to use.

PCR is quick but lacks sensitivity and those false negatives are a b***ard.

If you are based in the UK then use Mycoplasma Experience, a commercial company, COMPLETELY INDEPENDENT and therefore no conflict of interest........cheap at 85/cell line.

Kindest regards

Uncle Rhombus




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