Mycoplasma PCR tests for primates

B0009 - Ultrasensitive Mycoplasma screen by real time PCR. This screen detects but does not differentiate M. arginini, M. fermentans, M. hominis, M. hyorhinis, M. orale, M. pirum, M. salivarium, M. agassizii, M. cynos and others. This screen does not detect M. pneumoniae or M. pulmonis.

Mycoplasmas are gram-negative bacteria due to their absence of a cell wall. Because of this characteristic, these bacteria are not responsive to many common antibiotics such as penicillin or other beta-lactam antibiotics that target cell wall synthesis. There are more than 100 recognized species in the genus Mycoplasma. Some species cause disease in humans, such as M. pneumoniae which causes atypical pneumonia and other respiratory disorders, and M. genitalium which is linked to pelvic inflammatory diseases.

M. pneumoniae was first linked to respiratory infection more than one hundred years ago. M. pneumoniae can be transmitted through close contact via respiratory droplets. M. pneumoniae has a surface receptor which enables attachment to the host cell surface. This receptor can attach to a number of different cell types such as respiratory tract epithelia and red blood cells. At high concentrations, M. pneumoniae can inhibit ciliary action within the respiratory tract and cause cell necrosis. This damage is caused by cytotoxins from M. pneumoniae as well as indirectly from the host immune response.

Many other Mycoplasma species are found in research laboratories as contaminants in cell culture due to contamination from lab staff or contaminated cell culture medium ingredients. Contamination rates by Mycoplasma species in established cell cultures have been reported at 15-35%, with even higher occurrence cited in some populations. Mycoplasmas may induce cellular changes, including chromosome aberrations and changes in metabolism and cell growth. Severe Mycoplasma infections may even destroy a cell line. Therefore biological products derived from these cultures must be screened routinely to help ensure product stability and integrity. A number of techniques have been used to detect Mycoplasma, including DNA probe, enzyme immunoassays, PCR, plating on sensitive agar and staining with a DNA stain including DAPI or Hoechst. Among these, PCR is the most commonly used method due to its high sensitivity and specificity.

Mycoplasma pneumoniae on the other hand is difficult and slow to grow in culture media. However, molecular detection of M. pneumoniae by PCR is highly sensitive and specific, with fast turnaround time, so PCR is useful for detecting this species.

Utilities:

Help confirm the disease causing agent
Shorten the time required to confirm a clinical diagnosis of Mycoplasma infection.
Help ensure that animal populations are free of Mycoplasma species
Early prevention of spread of Mycoplasma species
Minimize personnel exposure to Mycoplasma species
Preservation of cell cultures
Safety monitoring of biological products

References:
Armstrong, S.E., Mariano, J.A. and Lundin, D.J. (2010) The scope of Mycoplasma contamination within the biopharmaceutical industry. Biologicals, 38:211-213.
Maczyñska, B., Matusiewicz, K., Chiciak, J., Stankiewicz, M., Sozañska, B. and Boznañski, A. (2002) Comparison of detectability of Mycoplasma pneumoniae infections in children, using PCR-test and serological methods: indirect immunofluorescence and immunoenzymatic assay. Clin. Microbiol. and Infect. 8 (Suppl. 1): P1346.

Specimen requirements: 0.2 ml whole blood in EDTA (purple top) tube, or 0.2 ml serum, CSF or bronchial lavage, or 0.2 ml fresh, frozen or fixed tissue, or nasopharyngeal swab, throat swab or vaginal swab, or 0.2 ml culture.

Contact Zoologix if advice is needed to determine an appropriate specimen type for a specific diagnostic application. For specimen types not listed here, please contact Zoologix to confirm specimen acceptability and shipping instructions.

For all specimen types, if there will be a delay in shipping, or during very warm weather, refrigerate specimens until shipped and ship with a cold pack unless more stringent shipping requirements are specified. Frozen specimens should be shipped so as to remain frozen in transit. See shipping instructions for more information.

Turnaround time: 2 business days

Methodology:
B0002 - Qualitative PCR
B0009 - Qualitative real time PCR

Normal range: Nondetected