AUTHOR=Ye Yin-Bo , Yang Jiang-Hua , Li Dong-Liang , Hao Li-Hua , Zhang Zhao , Mei Si-Yao , Zhang Huan , Du Fang-Yuan , Yv Li-Hui , Liu Bao-Shan , Chen Ze-Liang 

TITLE=A specific reverse complement sequence for distinguishing Brucella canis from other Brucella species

JOURNAL=Frontiers in Veterinary Science

VOLUME=9

YEAR=2022

URL=https://www.frontiersin.org/journals/veterinary-science/articles/10.3389/fvets.2022.983482

DOI=10.3389/fvets.2022.983482

ISSN=2297-1769

ABSTRACT=<p>Canine brucellosis is primarily caused by <italic>Brucella canis</italic>, but other <italic>Brucella</italic> species can also cause the disease. Identifying sequences specific to <italic>B. canis</italic> and establishing PCR assays that can distinguish between <italic>B. canis</italic> and other <italic>Brucella</italic> species is essential to determine the etiology of canine brucellosis and the source of infection and to achieve effective control. We analyzed the gaps and SNPs of genomes I and II from <italic>B. canis</italic> strain RM6/66 and <italic>B. melitensis</italic> strain 16M using the Mauve genome alignment software, and the specificity of each of these differential regions was analyzed by BLAST. A 132 bp specific sequence was found between the DK60_915 (glycosyl hydrolase 108 family protein) and DK60_917 (aldose 1-epimerase) loci in <italic>B. canis</italic> chromosome 1. Further comparative analysis revealed that this is a reverse complement sequence between <italic>B. canis</italic> and other <italic>Brucella</italic> species. Then, three primers were designed based on the sequence that could detect <italic>B. canis</italic> with a 310 bp amplification product or other <italic>Brucella</italic> species with a 413 bp product. The PCR based on these primers had reasonable specificity and a sensitivity of 100 copies of <italic>Brucella</italic> DNA. The detection results for the blood samples of the aborted dogs showed a favorable accordance with the Bruce-ladder multiplex PCR assay. In conclusion, we found a specific reverse complement sequence between <italic>B. canis</italic> and other <italic>Brucella</italic> and developed a PCR method that allows a more comprehensive identification of the pathogen involved in canine brucellosis. These findings provide an effective means for preventing and controlling brucellosis.</p>