Rapid detection of Mycoplasma bovis by real-time Loop-mediated isothermal amplification

Authors

  • Yan Pan
  • Ying Ming Wei
  • Garmendia AE
  • Hao Peng
  • Shi Wen Feng
  • Li Tao
  • Jun Li

Keywords:

Mycoplasma bovis, LAMP, OPPD/F gene

Abstract

Mycoplasma bovis (M. bovis) is one of the most significant pathogens in cattle. The aim of this study was to develop
a rapid real-time LAMP assay, specific for M. bovis with designed primers targeting the OPPD/F gene. The assay was
tested for 1 h at 63℃. The results could be visualized by the addition of calcein dye. Real-time LAMP was specific
exclusively for M. bovis without cross-reactivity with other bacteria. The sensitivity of the real-time LAMP was
determined to be 2.29×10-3ng/μL of DNA and was 100 times higher than conventional PCR. Lamp assay was conducted
on 65 clinical samples to test its usefulness and reliability and compared to the PCR assay results. A concordance of
100% was observed between the two assays. This real-time LAMP assay can be applied by diagnostic laboratories and
for field detection of M. bovis infection.

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Published

2020-09-17

How to Cite

Pan, Y. ., Wei, Y. M. ., AE, G. ., Peng, H. ., Feng, S. W. ., Tao, L. ., & Li, J. . (2020). Rapid detection of Mycoplasma bovis by real-time Loop-mediated isothermal amplification. The Thai Journal of Veterinary Medicine, 50(3), 363–369. Retrieved from https://he01.tci-thaijo.org/index.php/tjvm/article/view/245846

Issue

Vol. 50 No. 3 (2020): September

Section

Original Articles