Mycoplasma Test

 

The offered service is designed to detect contaminations with

  • Mycoplasma (Identification of 14 specific strains: 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)

Detection of more than 100 additional Mycoplasma strains by proprietary universal probes. This service detects all Mollicutes mentioned in the European Pharmacopoeia 2.6.7. Mycoplasma specification enables tracing of Mycoplasma contamination sources.

  

Test specifications

Mycoplasma testing is done with specific primer sequences directed to the highly conserved 16S rRNA coding region in the Mycoplasma genome and subsequent hybridisation using specific oligonucleotide probes. For internal control of DNA quality the presence of the PolA gene is tested as a reference. Several positive and negative controls are included to test PCR performance.

 

To be done by the user

  • Cell line lysate preparation according to the lysate protocol
    • required amount: >15µL
    • tubes: 1.5 ml SafeLock tubes (please do not use screw caps and Parafilm)
  • Complete the
  • Label sample tubes with:
    • the specific sample name as provided in the confirmation Email
  • Ship samples in a padded envelope WITH a copy of the confirmation Email to:

 Multiplexion, c/o DKFZ, Dept. F020, INF 242, D-69120 Heidelberg, Germany

 

To be done by Multiplexion

  • Mycoplasma testing
  • Transfer of result report to customer by Email (the report can be used as certificate for granting agencies and journals)

  

Turnaround Time

Samples submitted are run in even calendar weeks, with results emailed usually by end of the week.

  • Customers from Heidelberg should drop off samples in the building INF242 between 9AM and 12AM on Monday for inclusion in that week's processing schedule.
  • External customers should ship samples to the lab to be received by Monday for inclusion in that week's processing schedule. Samples received Tuesday through Friday of any given week will be held and processed the following testing week.

 

Special Preparation for Cell Culture Samples

Elimination of antibiotics is not necessary, however, to maximize Mycoplasma test sensitivity, cell lines should be pre-cultured in the absence of antibiotics for several days.

 

Mycoplasma elimination

  • Different contaminations require different actions. An overview can be found here.
  • You may use the following protocol to eliminate Mycoplasma from your precious cell lines. The protocol was kindly provided by U. Koch (DKFZ, Heidelberg).
  • Problems with contaminations? Our check list for sterile work may help.

 

Please note: This service is FOR RESEARCH USE ONLY. Do not use in diagnostic procedures.

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Important

No testing will be performed in week 32 of 2017. All samples will be collected during that time and tested in week 34.

Lysate preparation protocol

Download the detailed lysate
preparation protocol.

 

Technical Note

Analytical sensitivity of the
Multiplex cell Contamination Test.

 

Check list for sterile work

Download Multiplexion´s
check list for sterile work.

 

Mycoplasma

The PCR primer sets used in McCT are directed to the highly conserved 16S rRNA coding region in the Mycoplasma genome. This allows for amplification of all Mycoplasma and Acholeplasma species usually encountered as contaminants in cell cultures. Moreover, the McCT assay determines the genotype of Mycoplasma and Acholeplasma species by specific hybridisation, such as M. arginini, M. fermentans, M. orale, M. hyorhinis, M. hominis, M. salivarium, M. synoviae, M. pirum, M. gallisepticum, M. pneumoniae, M. yeatsii, and Acholeplasma laidlawii.

Mycoplasma specification enables the identification of contamination sources:

  mycoplasma species