The invention discloses primers, probes and a kit for rapidly detecting
mycoplasma bovis on site. The kit comprises a
forward primer sequence shown as SEQ No.1, a
reverse primer sequence shown as SEQ No.2, and a probe sequence shown as SEQ No.3, wherein the 5'-terminal of the
reverse primer sequence is labeled by
biotin; the 5'-terminal of the probe sequence is labeled by FAM, dSpacer is arranged at the position away from the 5'-terminal by 30 basic groups, and the 3'-terminal is blocked by C3-spacer. The primers, the probe
assembly and the kit for detecting the
mycoplasma bovis RPA-nfo are high in sensitivity and strong in specificity, the
mycoplasma bovis
DNA of 10 copies / reaction can be detected at the minimum, and the primers and the probes for detecting RPA-nfo respectively have no cross reaction with other mycoplasmas, as well as the
bacteria including
pasteurella multocida, mannheimia haemolytica, arcanobacterium pyogenes, histophilus somni, and
streptococcus pneumoniae.