A new Brucella canis species-specific PCR assay for the diagnosis of canine brucellosis
A new Brucella canis species-specific PCR assay for the diagnosis of canine brucellosis
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DOI：
10.1016/j.cimid.2014.07.003
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发表时间：
2014-09-04
影响因子：
2
通讯作者：
Her, Moon
中科院分区：
文献类型：
--
作者：
Kang, Sung-Il;Lee, Sang-Tun;Her, Moon
布鲁氏菌病是一种人畜共患疾病，由动物传播给人类，开发快速、准确和广泛可用的鉴定方法对诊断这种疾病至关重要。本研究建立了一种新的布鲁氏菌种特异性（BcSS）PCR检测方法，并对其特异性和敏感性进行了评价。根据B的II号染色体BCAN_B0548-0549区域设计特异性PCR引物。犬。B.犬种布鲁氏菌包括300 bp产物的扩增，该产物在其他布鲁氏菌种属或遗传学或血清学相关细菌上未发现。BcSS-PCR检测的检出限为6 pg/μ l（DNA稀释法），或从实验接种B的全血中分离的血沉棕黄层中的3 × 10（3）菌落形成单位（CFU）。罐。在该PCR测定中使用血沉棕黄层导致对B的灵敏度提高约100倍。与直接从全血中检测相比，这是第一个报告的种特异性PCR检测B。新的检测方法将为B的诊断提供有价值的工具。犬感染。(C)2014爱思唯尔有限公司版权所有。
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. (C) 2014 Elsevier Ltd. All rights reserved.