Development of a Single PCR Assay for Detection of Four Kinds of Brucella in Dogs
In order to establish a single PCR assay appropriate for detection of brucella in dogs,one pair of primers were designed through comparison of genomic DNA sequences between rough phenotype(B. canis)and smooth phenotype(B. melitensis,B. abortus and B. suis)strains,after optimization of reaction conditions,a PCR assay was established for preliminary detection of the four kinds of brucella,then its specificity and sensitivity were evaluated,and 20 Brucella-seropositive blood samples from dogs were tested clinically. The results showed that,for B. melitensis,B. abortus and B. suis,the fragment with the length of 717 bp could be amplified by the established method,and for B. canis,the fragment with the length of 366 bp could be amplified;the lowest genomic DNA concentrations of B. canis,B. melitensis,B. suis and B. abortus were 5.07×10-2,6.20×10-2,7.80×10-3 and 5.50×10-2 ng/μL,respectively;three positive samples for B. canis were detected out in 20 blood samples,which was consistent with the PCR results of using GB/T 18646—2018 primers. As a conclusion,the established assay was appropriate for detection of brucella in dogs in field veterinary laboratories due to its advantages of simplicity,specificity and sensitivity.
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