A new Brucella canis species-specific PCR assay for the diagnosis of canine brucellosis |
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| Authors: | Sung-Il Kang Sang-Eun Lee Ji-Yeon Kim Kichan Lee Jong-Wan Kim Hyang-Keun Lee So-Ra Sung Young-Ran Heo Suk Chan Jung Moon Her |
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| Institution: | 1. The OIE Reference Laboratory for Brucellosis, Bacterial Disease Division, Animal and Plant Quarantine Agency, 480 Anyang, Gyeonggi-do 430-757, South Korea;2. Division of Malaria & Parasitic Diseases, Korea National Institute of Health, Korea Centers for Disease Control & Prevention, Chengwon-gun, South Korea;3. Department of Food and Nutrition, Chonnam National University, Yongbongdong, Gwanju, South Korea |
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| Abstract: | 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 canis 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/μl by DNA dilution, or 3 × 103 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. |
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| Keywords: | Brucella canis Buffy coat Diagnostics Species-specific PCR assay Zoonosis |
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