title: A new Brucella canis species-specific PCR assay for the diagnosis of canine brucellosis.
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 6pg/μl by DNA dilution, or 3×10(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.
authors.affiliation: The OIE Reference Laboratory for Brucellosis, Bacterial Disease Division, Animal and Plant Quarantine Agency, 480 Anyang, Gyeonggi-do 430-757, South Korea.
authors.forename: Sung-Il
authors.initials: SI
authors.lastname: Kang
authors.affiliation: Division of Malaria & Parasitic Diseases, Korea National Institute of Health, Korea Centers for Disease Control & Prevention, Chengwon-gun, South Korea.
authors.forename: Sang-Eun
authors.initials: SE
authors.lastname: Lee
authors.affiliation: The OIE Reference Laboratory for Brucellosis, Bacterial Disease Division, Animal and Plant Quarantine Agency, 480 Anyang, Gyeonggi-do 430-757, South Korea.
authors.forename: Ji-Yeon
authors.initials: JY
authors.lastname: Kim
authors.affiliation: The OIE Reference Laboratory for Brucellosis, Bacterial Disease Division, Animal and Plant Quarantine Agency, 480 Anyang, Gyeonggi-do 430-757, South Korea.
authors.forename: Kichan
authors.initials: K
authors.lastname: Lee
authors.affiliation: The OIE Reference Laboratory for Brucellosis, Bacterial Disease Division, Animal and Plant Quarantine Agency, 480 Anyang, Gyeonggi-do 430-757, South Korea.
authors.forename: Jong-Wan
authors.initials: JW
authors.lastname: Kim
authors.affiliation: The OIE Reference Laboratory for Brucellosis, Bacterial Disease Division, Animal and Plant Quarantine Agency, 480 Anyang, Gyeonggi-do 430-757, South Korea.
authors.forename: Hyang-Keun
authors.initials: HK
authors.lastname: Lee
authors.affiliation: The OIE Reference Laboratory for Brucellosis, Bacterial Disease Division, Animal and Plant Quarantine Agency, 480 Anyang, Gyeonggi-do 430-757, South Korea.
authors.forename: So-Ra
authors.initials: SR
authors.lastname: Sung
authors.affiliation: Department of Food and Nutrition, Chonnam National University, Yongbongdong, Gwanju, South Korea.
authors.forename: Young-Ran
authors.initials: YR
authors.lastname: Heo
authors.affiliation: The OIE Reference Laboratory for Brucellosis, Bacterial Disease Division, Animal and Plant Quarantine Agency, 480 Anyang, Gyeonggi-do 430-757, South Korea.
authors.forename: Suk Chan
authors.initials: SC
authors.lastname: Jung
authors.affiliation: The OIE Reference Laboratory for Brucellosis, Bacterial Disease Division, Animal and Plant Quarantine Agency, 480 Anyang, Gyeonggi-do 430-757, South Korea. Electronic address: herm@korea.kr.
authors.forename: Moon
authors.initials: M
authors.lastname: Her
mesh_terms: D000818:Animals; D041002:Brucella canis / Q000235:genetics*; D002006:Brucellosis / Q000662:veterinary*; D004283:Dog Diseases / Q000175:diagnosis* / Q000382:microbiology*; D004285:Dogs; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S; D012680:Sensitivity and Specificity
chemical_list: D012336:RNA, Ribosomal, 16S
keywords: Brucella canis; Buffy coat; Diagnostics; Species-specific PCR assay; Zoonosis