The invention provides real-time fluorescent quantitative PCR
nucleic acid sequences and a kit for detecting
Clostridium botulinum type A. The
nucleic acid sequences comprise an upstream primer A-F, adownstream primer A-R and a probe A-P. As the primers and the probe provided by the invention are used for detecting
Clostridium botulinum type A, detection accuracy reaches 100%; and when the primers and the probe are used for amplification of other 25 intestinal
pathogenic bacteria and common bacterial DNAs, no specific amplification curve appears, so the primers and the probe have good specificity. According to a
fluorescence quantitative PCR
standard curve constructed on the basis of a recombinant
plasmid standard substance, it is determined that the detection sensitivity to
Clostridium botulinum type A is 5.04*10<2> copies / [mu]l. In a fluorescent quantitative PCR detection
system, when the concentration of a recombinant
plasmid containing the A
toxin gene is in the range of 10<2>-10<9> copies / [mu]l, the Ct values of amplification reactions and template concentrations are in good
linear relationship. The real-time fluorescent quantitative PCR
nucleic acid sequences and the kit ofinvention have the characteristics of sensitive reaction, high specificity,
rapidity, convenience, high
throughput, etc., and provide an effective detection means for screening, large-scale quarantineand epidemiological investigation of
Clostridium botulinum type A.