Detection of mycoplasma contamination in cell cultures and bovine sera

Contamination of cell cultures and sera used for animal virus propagation with mycoplasmas represents a serious problem, especially in virology. Therefore specific control measures must be used. To achieve this we introduced PCR for the detection of mycoplasma species in cell cultures and compared its results with ELISA and microbiological culture. Seven mycoplasma species which are the most common contaminants of cell lines (Mycoplasma arginini, M. fermentans, M. hyorhinis, M. bovis, M. orale, M. hominis, and Acholeplasma laidlawii) were used to verify the method. Then we assessed five selected cell lines and three bovine sera by the PCR, ELISA and culture methods and compared the results. PCR was positive for all of the mycoplasma species tested. ELISA kit used (Mycoplasma detection kit, Roche, Germany) allowed detection of only four species of contaminating mycoplasmas (Acholeplasma laidlawii, Mycoplasma arginini, M. hyorhinis, and M. orale). All the methods detected contamination of the VERO and RK13 cell lines. The agents of contamination were determined by the species-specific ELISA kit as Mycoplasma arginini and M. orale, respectively. Other cell lines and sera tested were not contaminated with mycoplasma. The results confirmed that the PCR method used in the present study is a sensitive, fast and specific detection method of mycoplasma contaminations and is suitable for routine mycoplasma detection in cell cultures and bovine sera.