Detection Of The Pathogen Of Diarrhea In Fur Animals And Establishment Of A Triple PCR Method For Viral Diseases

In recent years,the mortality of fur animals caused by diarrhoeal diseases has gradually increased,causing huge economic losses to fur animal breeding industry.At present,there are some problems in Hebei Province,such as unclear types of pathogenic bacteria,low accuracy and high cost of related detection technology.Based on this,this study mainly carried out epidemiological investigation on pathogenic bacteria and viruses of fur animal diarrhea in Hebei Province,and established a multiplex PCR rapid detection method that can detect three common viral diseases at the same time,in order to provide reference for the rapid detection of fur animal diarrhea To provide the basis for diagnosis and prevention.The research results are as follows1.The detection rate of canine parvovirus in 198 samples of fur animals from Hebei Province was 40.40%.The detection rate of canine coronavirus and canine distemper virus was 0.51%and 4.04%,respectively.The detection rate of Escherichia coli and Salmonella was 10.61%and 5.05%respectively,which indicated that the prevention and control measures of fur animals should be strengthened.In addition,the results also showed that there was mixed infection caused by the two viruses in the collected samples.The mixed infection rate of canine parvovirus and canine coronavirus was 0.51%,and the mixed infection rate of canine parvovirus and canine coronavirus was 6.06%.2.The 80 canine parvovirus isolates were compared by BLSAT,and the homology analysis showed that all the detected viruses belonged to the major branch of canine parvovirus,which might belong to the branch of Chinese isolate mf996334.1.3.The specific fragments of CPV(700 bp),CCV(410 bp)and CDV(197 bp)were amplified by the designed primers.The gel was recovered and the positive recombinant plasmid was constructed.The established triple PCR method had good specificity for the detection of unrelated pathogens such as Escherichia coli,Salmonella,Klebsiella pneumoniae,Pseudomonas aeruginosa and Streptococcus.The lowest detectable copies of CPV,CDV and CCV were 1.56×10~9copies/UL,1.82×10 ~9and 1.30×10 ~9copies/UL respectively,which provided a technical means for the differential diagnosis of mixed infection caused by three kinds of fur animal diarrhea virus.