Detection of Mycoplasma

Mycoplasma agar base (80 ml) (Oxoid, Basingstoke, Hants, UK) add 2.8 g of agar base to 80 ml of water, mix and autoclave at 15 psi for 15 min. Prepare fresh as necessary. 

Dispense in universal containers in 10 ml aliquots. Heat to 56°C for 45 min and store at -30°C. 

Prepared medium must be used within 10 days. 

Autoclave agar at 15 psi for 15 min. Cool to 50°C and mix with the other constituents (which have been warmed to 50°C). 

Dispense 8 ml per 5 cm diameter Petri dish. Store at 4°C in sealed plastic bags. 

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GL34 Biologicals Mycoplasma Test for the detection of Mycoplasma contamination... 

Such a catabolic reaction is indeed excluded in 6 alkyl purine derivatives. The parent compound of this group, 6-methylpurine, is known for its cytotoxicity its libera tion from the 2 -deoxyribonucleoside by purine nucleo side phosphorylases is used for detection of mycoplasma in cell cultures.19 It is highly potent and toxic to nonproliferating and proliferating tumor cells. Recently, the use of cytotoxic bases liberated by purine nucleoside phosphorylases such as 6-methylpurine was proposed as a novel principle in the gene therapy of cancer.20... 

A range of assay techniques are available for the detection of mycoplasma contamination. These include staining, culture, DNA probes and co-cultivation. To... 

Note Both methods used for the detection of mycoplasma have the following general principles in common. The cells to be tested should, before testing, complete at least two passages in antibiotic-free media. Infection may be hidden by the presence of antibiotics. Cell cultures tested from frozen ampoules should undergo at least two passages because cryoprotectants may also mask infection. 

Figure 1.7.1 Detection of mycoplasma by PCR in experimentally infected Vero cells. Amplifications performed with the primer set mollil + moUi2a (lane 1) Vero cells infected with M. arginini (lane 2) Vero cells infected with M. fermentans (lane 3) Vero cells infected with M. hyorhinis (lane 4) Vero cells infected with M. orale (lane 5) Vero cells infected with A. laidlawiv, (lane 11) moUicute-free Vero cells (lane 13) negative control (distilled water). Amplifications performed with the primer set mollil + molli2b (lane 6) Vero cells infected with A. laidlawii (lane 7) Vero cells infected with M. arginini, (lane 8) Vero cells infected with M. orale-, (lane 9) Vero cells infected with M. fermentans (lane 10) Vero cells infected with M. hyorhinis (lane 12) mollicute-free Vero cells (lane 14) negative control.

Ponnamma, K.N., Rajeev, G. and Solomon, J.J. (1991). Detection of mycoplasma-like organisms in Proutista moesta (Westwood) a putative vector of yellow leaf disease of arecanut. Journal of Plants Crops, 19 63-65. 

Rocha, A.D.A., Okhi, S.T. and Hiruki, C. (1986). Detection of mycoplasma like organisms in situ by indirect immunofluorescence microscopy. Phytopathology, 76 864-868. 

Detection of Mycoplasma Alison Stacey and Glyn Stacey... 

Testing of cell cultures for the presence of key adventitious agents should be routine in any tissue culture facility. Altough bacterial and fungal contaminations can be detected by microscopic and sometimes by macroscopic examination, the detection of mycoplasma and virus contaminants require the use of specific test procedures, including isolation by culture, PCR methods, electron microscopy, and analysis of cytopathic effects. 

Comparison of Different Techniques Used for the Detection of Mycoplasma Infection... 

Bl. Bernet, C., Garret, M., de Barbeyrac, B Bebear, C., and Bonnet, J. Detection of Mycoplasma pneumoniae by using the polymerase chain reaction. J. Clin. Microbiol. 27,2492-2496 (1989). 

Jl. Jensen, J. S., Uldum, S. A., Sonderg ard-Andersen, J., Vuust, J., and Lind, K., Polymerase chain reaction for detection of Mycoplasma genilalium in clinical samples. J. Clin. Micro-biol.29, 46-50 (1991). 

Another approach towards human glyco-sylation is followed by the utilization of a humanized mouse cell line, a human/ mouse heterohybridoma presented by Dr. Volker Sandig from ProBioGen. I have known Volker for a couple of years, since when I invented a real-time PGR test kit for the detection of mycoplasma contamination in pharmaceutical products. Using an internal standard we developed this method for rapid in-process (IPG) control for production of biopharmaceuticals and ProBioGen was one of the partners participating in the validation of the system as they wanted to use it for rapid quality analysis of their designer cell lines. As we have learned from previous examples, the development of mammahan super-producer cells from CHO (or also from the mouse myeloma cell line, NSO) starter cell... 

Cheng HS, Shen CW, Wang SR (2007) Effect of storage conditions on detection of mycoplasma in hiopharmaceutical products. In Vitro Cell Dev Biol Anim 43 113-119... 

Xiao JH, Liu Y, Wang MG, Jiang CH, You XX, Zhu CM. Detection of Mycoplasma pneumoniae PI subtype variations by denaturing gradient gel electrophoresis. Diagn Micr Infec Dis. 2014 78(l) 24-8. 

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