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Mycoplasma Quality Control

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Prevalence and effect of mycoplasma contamination

Mycoplasma contamination affects roughly 15-35% of continuous cell cultures, resulting in a number of deleterious effects, including:

  • Inhibition of cell metabolism
  • Induction of chromosomal abnormalities
  • Disruption of DNA and RNA synthesis
  • Changes in virus and antibody production
  • Altered gene expression in cells
  • Interference with growth rate of cells
  • Depletion of arginine
  • Change in pH
  • Activation of B cells
  • Decreased transfection rates
Importance of routine testing

These adverse effects can lead to the misinterpretation of results and compromise the validity of data generated for research and/or development projects.

Because mycoplasma can affect the phenotypic and functional characteristics of cells in vitro, the implementation of early detection methods that are rapid and sensitive is an important step towards preventing the deleterious and costly effects of mycoplasma on research and development projects.

The most frequently used detection methods include direct culture, Hoechst DNA staining, and PCR- when validated as a comparable method of detection.

Mycoplasma Quality Control

Assay development and validation

Developing and implementing a novel PCR-based mycoplasma detection system can be challenging and time consuming with regard to sample preparation and validation of the system to ensure equivalency or superiority to conventional test methods. To meet this need, ATCC has developed titered mycoplasma reference standards for comparing PCR and culture-based detection methods, and quantitative mycoplasma DNA certified reference materials for use as external controls in inclusivity/ exclusivity testing and establishing limits of detection. These products are rigorously authenticated and characterized by ATCC ISO 9001 certified laboratories, and represent a unique collection of species that are commonly associated with 95% of all mycoplasma contamination in cell culture.

ATCC Titered Mycoplasma Reference Strains Panel (ATCC® No. MP-7™)

A panel of 10 titered mycoplasma reference strains commonly associated with cell culture contamination. Each strain is prepared with a low genome copy to colony forming unit ratio, which is ideal for use in the development and validation of PCR-based methods of detection.

ATCC® No. Organism Designation Source
23206-TTR™ Acholeplasma laidlawii PG8 Sewage
23838-TTR™ Mycoplasma arginini G230 Mouse brain experimentally infected with scrapies
19989-TTR™ Mycoplasma fermentans PG18 Ulcerative balanitis
19610-TTR™ Mycoplasma gallisepticum Suspension of tracheal and air sac tissues of chickens with chronic respiratory disease
27545-TTR™ Mycoplasma hominis Human blood
17981-TTR™ Mycoplasma hyorhinis BTS-7 Nasal cavity of a pig
23714-TTR™ Mycoplasma orale CH 19299 Human – oropharynx of a child
15531-TTR™ Mycoplasma pneumoniae FH strain of Eaton Agent Human – atypical pneumonia
23064-TTR™ Mycoplasma salivarium Saliva
25204-TTR™ Mycoplasma synoviae WVU 1853 Hock joint of a chicken
ATCC Quantitative Mycoplasma DNA Certified Reference Materials

These certified reference materials are derived from the strains represented in the ATCC Titered Mycoplasma Reference Strains Panel (ATCC® No. MP-7™). Each preparation is produced under an ISO Guide 17034 accredited process to confirm identity, well-define characteristics, and an established chain of custody. These tools are ideal for use in establishing limits of detection, inclusivity/ exclusivity testing, and validating or comparing test methods.

ATCC® No. Organism Designation Specification Range
qCRM-23206D™ Acholeplasma laidlawii PG8 1x106 – 1x107 genome copies/µL
qCRM-23838D™ Mycoplasma arginini G230 1x106 – 1x107 genome copies/µL
qCRM-19989D™ Mycoplasma fermentans PG18 1x106 – 1x107 genome copies/µL
qCRM-19610D™ Mycoplasma gallisepticum 1x106 – 1x107 genome copies/µL
qCRM-27545D™ Mycoplasma hominis 1x106 – 1x107 genome copies/µL
qCRM-17981D™ Mycoplasma hyorhinis BTS-7 1x106 – 1x107 genome copies/µL
qCRM-23714D™ Mycoplasma orale CH 19299 1x106 – 1x107 genome copies/µL
qCRM-15531D™ Mycoplasma pneumoniae FH strain of Eaton Agent 1x106 – 1x107 genome copies/µL
qCRM-23064D™ Mycoplasma salivarium 1x106 – 1x107 genome copies/µL
qCRM-25204D™ Mycoplasma synoviae WVU 1853 1x106 – 1x107 genome copies/µL

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