Acta Vet. Brno 2020, 89: 317-332
Molecular genetic techniques and oligonucleotides for mycoplasma identification – a review
Control of distribution of mycoplasmal infections in cattle herds is essential in the majority of countries world-wide. Various PCR procedures are available to detect mycoplasmas in cell cultures and bovine mycoplasma in different types of samples. We reviewed some common PCR techniques and specific primers targeted to different bacterial genetic regions of mycoplasma. Several researchers used the same PCR approach and Mycoplasma spp. as a target but their results could not be compared because different primer pairs were used. These methods and primers were first developed to identify mycoplasma species that contaminate animal cell cultures, and then were used by other researchers to differentiate mycoplasmas as a cow infecting agent. Our analysis of the specificity of these primer pairs to nucleotide sequences of five Mycoplasma spp. showed that oligonucleotides have less specificity to them. Numerous commercially available PCR kits are applicable to find mycoplasma contamination in cell cultures and fewer of them can be used in veterinary diagnostics. Although serological and culture techniques are still used, it is necessary to develop a new multiplex PCR technique with a more specific primer set especially in agrarian countries.
Keywords
PCR, species-specific primers, sequencing, nested PCR, real-time PCR, cow disease.
Funding
The study was supported by the Ministry of Agriculture of the Republic of Kazakhstan and the Kazakh Scientific Research Veterinary Institute as part of targeted funding for the program “Scientific support for veterinary welfare and food safety” 267/101/156.
