Mycoplasma Detection Methods using PCR

Five species are considered to be responsible for more than 95% of cell culture contaminations M. arginini, M. fermentans, M. orale, M. hyorhinis and A. laidlawii. 

Many different techniques, such as bacteriological culture, DNA staining using fluorochrome and immunological or biochemical methods, are available to detect mycoplasma contamination (see section 1.6). However, none seem to be fully efficient, so a combination of different methods is often necessary. Molecular tools such as hybridization using rDNA gene probes or polymerase chain reaction (PCR) have been developed over the past few years. Several studies using 16S rDNA-based PCR concluded that PCR seems to be a very convenient method for routine detection of cell culture contaminations (Spaepen, 1992 Teyssou, 1993 van Kuppeveld, 1994). 

A UV light is damaging to eyes. Always wear protective glasses. 

A photograph of the gel can be taken using a bellows-type camera equipped with a Polaroid MP4 system using 3000 iso-Polaroid 667 film. An orange filter is necessary to obtain a good film image. The time of exposure required is 0.5-1 s with a fully opened diaphragm. 

Cell and Tissue Culture Laboratory Procedures in Biotechnology, edited by A. Doyle and J.B. Griffiths. 1998 John Wiley Sons Ltd. 

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