An improved loop-mediated isothermal amplification assay for the detection of Mycoplasma bovis.
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.
Department of Veterinary Medical Science, Faculty of Agriculture, University of Miyazaki, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan.
Yumiko
Y
Higa
Ryoko
R
Uemura
Wataru
W
Yamazaki
Shinya
S
Goto
Yoshitaka
Y
Goto
Masuo
M
Sueyoshi
D000818:Animals; D048090:Bovine Respiratory Disease Complex / Q000175:diagnosis* / Q000382:microbiology; D002417:Cattle; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000662:veterinary*; D045682:Mycoplasma bovis; D021141:Nucleic Acid Amplification Techniques / Q000379:methods / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity