Abstract

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.

Highlights

  • Control of distribution of mycoplasmal infections in cattle herds is essential in the majority of countries world-wide

  • The results of this study showed that milk samples from a bulk tank with a negative result of microbiological assessment had a positive test for the presence of M. bovigenitalium in the SYBR-PCR assays

  • In their research for rt-PCR identification of M. bovigenitalium and M. californicum, Parker et al (2017) applied DNA probes and primers MbvgF and MbvgR, McalF and McalR described by Boonyayatra et al (2012) and MbovP, MbovF and MbovR oligonucleotides specific to M. bovis uvrC gene and developed by Clothier et al (2010)

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Summary

Introduction

Control of distribution of mycoplasmal infections in cattle herds is essential in the majority of countries world-wide. For the detection and discrimination between different mycoplasma species as contaminants in cell cultures, Harasawa et al (1993) have developed a nested PCR method with two pairs of primers necessary to amplify a heterogeneous 16S-23S rRNA spacer region.

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