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A novel polymerase chain reaction assay for the detection of seven Mycoplasma species of cattle origin

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Abstract

The study aimed to develop a pair of polymerase chain reaction primers for detecting ruminant mycoplasma pathogens. We designed a set of primers based on the most similar sequences within 16 S rRNA regions of seven Mycoplasma spp. These primers have high sensitivity for detecting Mycoplasma dispar, M. arginine, M. canadense, M. bovis, M. alkalescens, M. californicum, and M. bovigenitalium within the annealing temperature range of 46 to 48 °C. The minimum amount of DNA that can be detected using the protocol is 250 ng, which is equivalent to 2,000 colony-forming units per mL. The primers can detect mycoplasma from DNA extracted directly from milk samples. The common bovine mastitis pathogens of Staphylococcus aureus coagulase-negative staphylococci, Escherichia coli, Streptococcus uberis, Klebsiella pneumonia, and Kocuria rosea were not detected by the primers. We believe the high sensitivity and specificity of these primers make them useful for detecting infection with seven Mycoplasma species in ruminants, allowing the primers to be used in clinical settings.

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Acknowledgements

The authors would like to thank all members of the Extension Center and all members of the laboratory managed by Dr. Nagahata and Dr. Higuchi at the Rakuno Gakuen University. The authors also thank the Rakuno Gakuen University International Guest Researcher Program for its support. This manuscript was edited by Wallace Academic Editing.

Funding

This study was funded by the Council of Agriculture, Executive Yuan of the Republic of China, Taiwan. No. 110AS-5.1.2-BQ-B1 and Rakuno Gakuen University International Guest Researcher Program in 2017.

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Authors and Affiliations

  1. Department of Veterinary Medicine, National Chiayi University, National Chiayi University, 60004, Chiayi, Taiwan

    Jyhmirn Lai & Hengching Lin

  2. Animal Health Unit, Department of Health and Environmental Sciences, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, Japan

    Satoshi Gondaira, Hidetoshi Higuchi & Hajime Nagahata

  3. Department of Veterinary Associated Science, Faculty of Veterinary Medicine, Okayama University of Science, Ikoinooka 1-3, 794-8555, Imabari, Ehime, Japan

    Hajime Nagahata

  4. Department of Early Childhood Educare, Tainan University of Technology, 710302, Tainan City, Taiwan

    Peichuan Hsu

Authors
  1. Jyhmirn Lai
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  2. Hengching Lin
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  3. Peichuan Hsu
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  4. Satoshi Gondaira
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  5. Hidetoshi Higuchi
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  6. Hajime Nagahata
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Contributions

JML and HN conceived and designed the research. JML, SG, and HH conducted experiments. JML, HCL, and HPH contributed new reagents or analytical tools. JML, HPH, and HCL analyzed the data. JML wrote the manuscript. All authors read and approved the manuscript. JML and HN managed the funding.

Corresponding author

Correspondence to Jyhmirn Lai.

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None of the authors have any conflicts of interest to declare.

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This article does not contain any studies with animals performed by any of the authors.

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This material is the authors’ own original work, which has not been previously published elsewhere. The paper is not currently being considered for publication elsewhere. The paper reflects the authors’ own research and analysis in a truthful and complete manner. The paper properly credits the meaningful contributions of co-authors and co-researchers. The results are appropriately placed in the context of prior and existing research. All sources used are properly disclosed. All authors have been personally and actively involved in substantial work leading to the paper, and will take public responsibility for its content. All authors agree to publish the manuscript in this journal.

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Lai, J., Lin, H., Hsu, P. et al. A novel polymerase chain reaction assay for the detection of seven Mycoplasma species of cattle origin. World J Microbiol Biotechnol 38, 128 (2022). https://doi.org/10.1007/s11274-022-03312-6

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  • DOI: https://doi.org/10.1007/s11274-022-03312-6

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