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Easy outsourcing of mycoplasma testing in any cell line
MYCOPLASMACHECK is a qPCR-based service for detection of mycoplasma in cell cultures. As contamination of mycoplasma alters cell-line properties, their unnoticed presence often leads to non-reproducible results. Regular testing with MYCOPLASMACHECK serves as a rapid, accurate and objective means of checking cells for mycoplasma contamination, ensuring cell lines are kept reliably clean.
Highlights of MYCOPLASMACHECK
- Standardised qPCR test for mycoplasma performed under ISO17025 accreditation
- High specificity and sensitivity guaranteed by use of internal controls
- Data provided online in 48 hours
- Test history report for select periods of time can be compiled online at any time
Full-service package offered by MYCOPLASMACHECK, including
- Detection of all mycoplasma species listed in the European Pharmacopoeia 2.6.7: M. arginini, M. fermentans, M. orale, M. hyorhinis, M. hominis, M. genitalium, M. salivarium, M. synoviae, M. pirum, M. gallisepticum, M. pneumoniae, M. yeatsii, Spiroplasma citri and Acholeplasma laidlawii
- Identification of more than 100 additional mollicutes
- Highly sensitive assay capable of detecting 1 international unit (IU) per PCR of the World Health Organization’s (WHO) international standard for mycoplasma DNA (Please see application note in "Files")
- Highly specific test that identifies ten copies of mycoplasma per reaction with no cross-reactivity to E. coli or eukaryotic DNA
- Use of positive and negative controls to monitor test performance
- Exclusively licensed from the German Cancer Research Center, (Deutsches Krebsforschungszentrum, DKFZ), Heidelberg, Germany
- Easy shipment via GATC Collection Points
Accepted starting material for MYCOPLASMACHECK
- 100-200 µL of heat-inactivated cell-culture supernatant (Please refer to our sample preparation protocol in "Files")
- Drop-off at any GATC Collection Point or ship separately
Technology used for MYCOPLASMACHECK
- Roche LightCycler 480 II
Deliverables of MYCOPLASMACHECK
- Data report indicating mycoplasma status, as well as any PCR inhibitors found in the sample
- The user can compile a PDF document online at any time to view the results from any specific testing period.
- PDF document serving as an independent certificate of results; suitable for direct submission to journals or granting agencies
Delivery time for MYCOPLASMACHECK
- Results delivered to your secure online myGATC account within 48 hours after sample receipt
Mycoplasma contamination adversely affects 35% of the world’s cell lines. As signs of mycoplasma are easy to miss, only regular monitoring of cell cultures can ensure that research results are valid and reproducible.
With its standardised workflows, GATC Biotech can rapidly and reliably detect all common mycoplasma species (Table 1).
Table 1. List of mycoplasma species frequently detected in cell cultures. These mycoplasma, all found in the European Pharmacopoeia 2.6.7., are readily detectable with MYCOPLASMACHECK.
As a qPCR-based assay, the technique is more accurate and sensitive than traditional microbial culture methods and bioluminescence-based methods for mycoplasma detection (Fig. 1).
Fig. 1. Comparison of three different methods for detection of mycoplasma in cell culture. The graph demonstrates the superiority of mycoplasma PCR over culture-based or bioluminescence-based (Mycoalert) techniques in analysing cell lines for mycoplasma (adapted from Cytotechnology (2014) 66:861–873)
Besides severely reducing the hands-on time involved in in-house testing, outsourcing the task of mycoplasma detection offers distinct benefits that cannot be matched by do-it-yourself kits (Fig. 2).
Fig. 2. Advantages of using MYCOPLASMACHECK for mycoplasma PCR detection over in-house PCR kits
1. What mycoplasma species are detected with MYCOPLASMACHECK?
MYCOPLASMACHECK can detect all mycoplasma species that contaminate cell cultures. All species, including M. arginini, M. fermentans, M. orale, M. hyorhinis, M. hominis, M. genitalium, M. salivarium, M. synoviae, M. pirum, M. gallisepticum, M. pneumoniae, M. yeatsii, Spiroplasma citri and Acholeplasma laidlawii, are listed in the European Pharmacopoeia 2.6.7. In silico analysis shows that more than 100 additional mollicutes can be detected with MYCOPLASMACHECK.
2. How sensitive is the MYCOPLASMACHECK PCR assay?
The qPCR test can detect a minimum of 10 mycoplasma copies per qPCR reaction.
3. What starting material is accepted for detection of mycoplasma?
We accept heat-inactivated cell-culture supernatant from any kind of eukaryotic cell line, as long as it is at or below risk group level 2.
4. How much material should I send for mycoplasma testing?
Only 100-200 µL of heat-inactivated cell-culture supernatant is required.
5. How should I prepare samples for MYCOPLASMACHECK?
A detailed protocol of how cells should be grown and how the cell-culture supernatant should be prepared is available in the Files tab.
6. When can I submit a sample for analysis?
You can submit a sample for mycoplasma detection at any time. We do not have pre-set testing dates and we analyse the samples for mycoplasma as soon as we receive them.
7. How soon do I receive the results?
The results of the test appear in your secure online myGATC account within 48 hours after sample receipt.
8. What do the results of MYCOPLASMACHECK look like?
The MYCOPLASMACHECK Data Report indicates the presence or absence of mycoplasma, as well as the presence or absence of PCR inhibitors. The customer is also given the exclusive opportunity to generate a PDF document containing only user-selected cell line results from any testing period. This customised PDF report can be directly used as a verification certificate for scientific journals and granting agencies. For a sample Data Report, please see "Files" section.
9. What happens if my culture is positive for mycoplasma contamination?
In the event of mycoplasma contamination, we recommend simply discarding the infected cell line. If the cell line is of significant value, then mycoplasma elimination procedures could be attempted. Numerous protocols and services for mycoplasma elimination are available and can be found online.
10. What could lead to PCR inhibition?
Most PCR inhibitors are found in cell cultures with >90% confluence or if suspension cells are present in the supernatant. Please refer to our "MYCOPLASMACHECK Sample Preparation" guide for instructions on how to best prepare your sample.
11. Where should I send my samples?
Please drop off your samples at any GATC Collection Point. Samples can also be shipped by mail:
GATC Biotech AG
European Custom Sequencing Centre