Early Detection of Brucella Canis via Quantitative Polymerase Chain Reaction Analysis
Early Detection of Brucella Canis via Quantitative Polymerase Chain Reaction Analysis
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DOI：
10.1111/zph.12041
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发表时间：
2014-02-01
影响因子：
2.4
通讯作者：
Petersen, C. A.
中科院分区：
文献类型：
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作者：
Kauffman, L. K.;Bjork, J. K.;Petersen, C. A.
犬布鲁氏菌病是一种可报告的人畜共患疾病，可通过接触受感染的尿液或其他泌尿生殖系统分泌物导致犬丧失生殖能力和人类感染。尽管许多地方要求对阳性犬进行检测和安乐死，但目前的诊断受到血清转化所需时间的限制，例如，是否存在犬b型特异性抗体。本研究的目的是确定犬布鲁氏菌特异性定量聚合酶链反应（qPCR）检测方法的诊断能力，以便在血清学阳性之前检测野外样本中的犬布鲁氏菌，从而更快地诊断和预防犬舍或家庭内的传播。在观察到至少一只狗的暗示临床症状和阳性血清学后，对两个犬舍（其中一个位于主人家中）进行了采样。所得标本经血清学和qPCR分析比较分析。通过qPCR对107只狗进行了犬B. canis感染分析：105只全血样本，65只阴道拭子样本，6只尿液样本，7只尸检时采集的泌尿生殖道组织样本。经qPCR检测发现45只犬感染犬布鲁氏菌病，其中血清阳性22只（48.89%）。有统计学意义（P = 0.0228）的qpcr阳性犬，5/25（20.00%）在首次血清学检测后30天内血清转化。与血清学相比，阴道拭子DNA qPCR分析的灵敏度为92.31%，特异性为51.92%，全血DNA qPCR分析的灵敏度为16.67%，特异性为100%。无创阴道拭子和尿液样本的犬B. canis外膜蛋白25 DNA qPCR可在检测抗体之前早期检测犬B. canis感染。该检测提供了一个关键的工具，以减少犬布鲁氏菌病的人畜共患传播，其相关的临床表现，以及情绪和经济影响。
Canine brucellosis is a reportable zoonotic disease that can lead to canine reproductive losses and human infection through contact with infected urine or other genitourinary secretions. Although many locations require testing and euthanasia of positive dogs, current diagnosis is limited by the time required for seroconversion, for example, presence of B. canis-specific antibodies. The goal of this study was to determine the diagnostic ability of Brucella canis-specific quantitative polymerase chain reaction (qPCR) assay to detect B. canis in field samples prior to serological positivity for faster diagnosis and prevention of transmission within kennels or in households. Two kennels, one of which was located in the owner's home, were sampled following observation of suggestive clinical signs and positive serology of at least one dog. Specimens obtained were comparatively analysed via serology and qPCR analysis. 107 dogs were analysed for B. canis infection via qPCR: 105 via whole-blood samples, 65 via vaginal swab, six via urine and seven via genitourinary tract tissue taken at necropsy. Forty-five dogs were found to be infected with canine brucellosis via qPCR, of which 22 (48.89%) were seropositive. A statistically significant number (P = 0.0228) of qPCR-positive dogs, 5/25 (20.00%), seroconverted within a 30-day interval after initial serologic testing. As compared to serology, qPCR analysis of DNA from vaginal swabs had a sensitivity of 92.31% and specificity of 51.92%, and qPCR analysis of DNA from wholeblood samples had a sensitivity of 16.67% and specificity of 100%. B. canis outer membrane protein 25 DNA qPCR from non-invasive vaginal swab and urine samples provided early detection of B. canis infection in dogs prior to detection of antibodies. This assay provides a critical tool to decrease zoonotic spread of canine brucellosis, its associated clinical presentation(s), and emotional and economic repercussions.