An improved loop-mediated isothermal amplification assay for the detection of            Mycoplasma bovis

Yumiko HIGA; Ryoko UEMURA; Wataru YAMAZAKI; Shinya GOTO; Yoshitaka GOTO; Masuo SUEYOSHI

doi:10.1292/jvms.15-0459

Bacteriology

An improved loop-mediated isothermal amplification assay for the detection of Mycoplasma bovis

Yumiko HIGA, Ryoko UEMURA, Wataru YAMAZAKI, Shinya GOTO, Yoshitaka GOTO, Masuo SUEYOSHI

Author information

Yumiko HIGA
Department of Veterinary Medical Science, Faculty of Agriculture, University of Miyazaki, 1–1 Gakuenkibanadai-nishi, Miyazaki 889–2192, Japan
Ryoko UEMURA
Department of Veterinary Medical Science, Faculty of Agriculture, University of Miyazaki, 1–1 Gakuenkibanadai-nishi, Miyazaki 889–2192, Japan
Center for Animal Disease Control, University of Miyazaki, 1–1 Gakuenkibanadai-nishi, Miyazaki 889–2192, Japan
Wataru YAMAZAKI
Department of Veterinary Medical Science, Faculty of Agriculture, University of Miyazaki, 1–1 Gakuenkibanadai-nishi, Miyazaki 889–2192, Japan
Center for Animal Disease Control, University of Miyazaki, 1–1 Gakuenkibanadai-nishi, Miyazaki 889–2192, Japan
Shinya GOTO
Department of Veterinary Medical Science, Faculty of Agriculture, University of Miyazaki, 1–1 Gakuenkibanadai-nishi, Miyazaki 889–2192, Japan
Yoshitaka GOTO
Department of Veterinary Medical Science, Faculty of Agriculture, University of Miyazaki, 1–1 Gakuenkibanadai-nishi, Miyazaki 889–2192, Japan
Center for Animal Disease Control, University of Miyazaki, 1–1 Gakuenkibanadai-nishi, Miyazaki 889–2192, Japan
Masuo SUEYOSHI
Department of Veterinary Medical Science, Faculty of Agriculture, University of Miyazaki, 1–1 Gakuenkibanadai-nishi, Miyazaki 889–2192, Japan
Center for Animal Disease Control, University of Miyazaki, 1–1 Gakuenkibanadai-nishi, Miyazaki 889–2192, Japan

Keywords: cattle, loop-mediated isothermal amplification (LAMP) assay, Mycoplasma bovis, respiratory disease

JOURNAL FREE ACCESS

2016 Volume 78 Issue 8 Pages 1343-1346

DOI https://doi.org/10.1292/jvms.15-0459

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Abstract

We improved a loop-mediated isothermal amplification (LAMP) assay permitting sensitive and rapid Mycoplasma bovis detection. A total of 55 bacterial strains were examined in this study, including 33 M. bovis strains, 14 non-M. bovis mycoplasmas and eight non-mycoplasma bacterial strains. M. bovis was successfully detected by the LAMP assay within 60 min without cross-reaction to any other bacteria. Furthermore, a total of 135 nasal swab samples were tested directly using our LAMP assays, the previously reported LAMP assay, conventional PCR assay without pre-culture and comparing standard culture methods. The improved LAMP assay showed sensitivity and specificity of 97.2% and 90.9%, respectively (with a kappa coefficient of 0.8231), and the sensitivity of our revised LAMP assay was increased compared to existing methods.

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