Kit for Mycoplasma detection by PCR
Posted 24 July 2002 - 01:56 AM
Posted 26 July 2002 - 09:32 AM
Posted 01 August 2002 - 08:54 AM
Posted 08 March 2005 - 03:02 AM
Very simple, reliable and inexpensive.
Posted 07 April 2010 - 09:01 AM
Posted 07 April 2010 - 05:07 PM
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.
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.