A new Brucella canis species-specific PCR assay for the diagnosis of canine brucellosis

作者：Kang Sung Il; Lee Sang Tun; Kim Ji Yeon; Lee Kichan; Kim Jong Wan; Lee Hyang Keun; Sung So Ra; Heo Young Ran; Jung Suk Chan; Her Moon^*

来源：Comparative Immunology, Microbiology and Infectious Diseases, 2014, 37(4): 237-241.

DOI：10.1016/j.cimid.2014.07.003

• Brucella canis
• Buffy coat
• Diagnostics
• Species-specific PCR assay
• Zoonosis

Brucellosis is a zoonotic disease that is transmitted from animals to humans, and the development of a rapid, accurate, and widely available identification method is essential for diagnosing this disease. In this study, we developed a new Brucella cants species-specific (BcSS) PCR assay and evaluated its specificity and sensitivity. A specific PCR primer set was designed based on the BCAN_B0548-0549 region in chromosome II of B. canis. The PCR detection for B. canis included amplification of a 300-bp product that is, not found on other Brucella species or, genetically or serologically related bacteria. The detection limit of BcSS-PCR assay was 6 pg/mu l by DNA dilution, or 3 x 10(3) colony-forming units (CFU) in the buffy coats separated from whole blood experimentally inoculated with B. canis. Using the buffy coat in this PCR assay resulted in approximately 100-times higher sensitivity for B. canis as compared to detect directly from whole blood. This is the first report of a species-specific PCR assay to detect B. canis, and the new assay will provide a valuable tool for the diagnosis of B. canis infection.

• 出版日期2014-9-4