A Guide to Mycoplasma Testing Services
Test information, sample requirements and shipping method
- Test Code / Protocol: 131
- Sample Options:
- Spent Medium: Provide 0.5 mL of antibiotic-free medium that has been on the cells for 3+ days. (Smaller volumes are acceptable; the DNA extraction protocol will be adjusted).
- Cell Vial: Send one frozen vial of cells.
- Any other material (biological, powder, gel, solid sample) can be shipped on ice packs or dry ice. The test requires 0.5 gram or 0.5 mL. Smaller volumes can be tested.
- Shipping: Transport on ice packs or dry ice. If the test article is stable at room temperature, you can ship at room temperature.
What is the turnaround time for the Mycoplasma testing by PCR?
- Delivery Time: 1–7 business days
- Digital Copy: Sent via PDF
Price per sample is $125. If you need a formal quote, please email Dr. Ahmed Kilani at akil@clongen.com to request a quote.
What is the report format for Mycoplasma testing?
You will receive a three-page report containing the sponsor’s information, materials and methods, and an electronically labeled gel image (including two Mycoplasma positive controls, one negative control, and the samples). The final results are clearly stated on the last page.
Do I need to notify the laboratory before shipping?
It is preferable to notify the lab by email ahead of shipping samples, especially if this is your first time to test with our lab.
What species of Mycoplasma are detected?
Use of test results:
For Research Use Only (RUO). Not intended for diagnostic, clinical, or therapeutic use, and not suitable for regulatory submissions, quality control release testing, or characterization of drug products under FDA scrutiny or cGMP guidelines.
Mycoplasma Elimination from contaminated cell lines
- 4 PCR Tests: One baseline screening, two interim monitoring tests during treatment, and one final clearance test five days post-treatment.
- Documentation: A detailed 5–7 page technical report including gel images and an official certificate of quality.
- Working Cell Bank: A minimum of 25 cryovials containing at least 1 million cells per vial, shipped securely on dry ice.
Turnaround time is 4 to 6 weeks, varying by cell line growth rates.
Cost: please email Dr. Ahmed Kilani with any questions and a formal price quote
Mycoplasma Testing Services
| Description | Protocol | Turnaround | Sample requirements |
|---|---|---|---|
| Mycoplasma species multiplex PCR detection - Non-GLP | 131 | 2-7 days | 0.5 mL or larger |
| Points to Consider 1993 Testing for Virus Stocks and Cell Lines (Mycoplasma Testing)-Cultivable and Non-Cultivable Mycoplasma Testing - GLP | CB120 | 4-5 weeks | 14 mL |
| Mycoplasma Elimination from Contaminated Cell Lines - Non GLP | CB124 | 4 - 6 weeks | 10 (7) cells |
| Mycoplasma Screen by Hoechst Staining (GLP) | CB115 | 1 - 2 weeks | 1 ml |
| Mycoplasma Screen by Hoechst Staining (Non-GLP) | CB114 | 1 - 2 weeks | 1 ml |
| Mycoplasma Testing by Direct Cultivation on enriched Mycoplasma Media-GLP | CB116 | 5 - 6 weeks | 5 ml |
Introduction and Background
Nocard and Roux reported the cultivation of the causative agent of Contagious Bovine Pleuro Pneumoniae (CBPP) in 1898, a disease that was widespread in cattle herds and was found to be caused by M. mycoides subsp. mycoides SC (small-colony type). In 1956, the first species of Mycoplasma were isolated in culture. The most commonly encountered species are Acholeplasma laidlawii, Mycoplasma arginini, M. orale, M. salivarium, M. fermentans, and M. hyorhinis. Species such as Ureaplasma urealyticum, M. pneumoniae and M. pirum are rarely present in cell cultures. It is anticipated that many of the cell lines created prior to 1990 were infected by Mycoplasma species that originated from contaminated animal serum and further disseminated by contaminated aerosols in the laboratory and laboratory technicians while they conducted their experiments.
A. laidlawii and M. arginini are of bovine origin; M. orale and M. salivarium are of a human origin (oropharynx) and M. hyorhinis originated from swine. It is hypothesized that presence of Mycoplasmas of swine origin in bovine serum is justified by the contamination of this product in mixed slaughterhouses.
It is estimated that as many as 30% of all cell lines generated prior to 1990 are contaminated with Mycoplasma species.
What is Mycoplasma doing to my cell lines??
Invisible in cell culture but can cause serious damage!
t is always a good practice to check your cell lines on a regular basis for Mycoplasma contamination. Mycoplasma contamination could be devastating to a cell culture facility and could result in heavy costs and wasted time if not detected early. At Clongen Laboratories, we offer several ways to test cell lines (or any sample source) for the presence of Mycoplasma, Ureaplasma and Acholeplasma contamination. One of our assays is a Cell-Based Assay where the test material (cell line or other) is inoculated onto a Vero cell monolayer known to be negative for the presence of Mycoplasma, incubated for a few days, fixed and stained with e Hoechst stain (DNA fluorochrome 33258). This fluorescence pattern of DNA in the cytoplasm of indicator cells indicates the infection status of the cells. Our laboratory scientists have many years of experience in evaluating microscopic fluorescence patterns and the assay is fairly sensitive (can detect 75-150 Mycoplasmas per ml). There are rare occasions where lysed cells or artifacts in the tested material can produce a false positive readout. We recommend a confirmatory assay for positive cultures other than the cell-based assay.
We also offer a highly sensitive Multiplex PCR (Polymerase Chain Reaction) assay for the detection of low copy number. The PCR assay is capable of detecting any of the 20 or more Mycoplasma and Acholeplasma species detected in cell culture.
At Clongen Labs, we offer aerobic and anaerobic cultivation of Mycoplasma on highly enriched special media used for the cultivation of Mycoplasma. Our assays are performed under GMP conditions and if requested, we offer GLP testing for Mycoplasma (21CFR 610.30) intended mainly for vaccine products. This assay detects both the cultivable and non-cultivable Mycoplasmas using the cell-culture based technique and the enriched microbiological medium to attempt to grow Mycoplasmas. We offer Mycoplasma GLP testing (9CFR113.28) for cultivable Mycoplasmas in enriched medium and the Points to Consider 1993 testing for Virus Stocks using cell-based and enriched medium growth. Protocols will be provided to clients upon request.