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骆驼血清中布鲁氏菌属检测诊断试验的比较。

Comparison of diagnostic tests for the detection of Brucella spp. in camel sera.

作者信息

Gwida Mayada M, El-Gohary Adel H, Melzer Falk, Tomaso Herbert, Rösler Uwe, Wernery Ulrich, Wernery Renate, Elschner Mandy C, Khan Iahtasham, Eickhoff Meike, Schöner Daniel, Neubauer Heinrich

机构信息

Friedrich-Loeffler-Institut, Institute for Bacterial Infections and Zoonoses, Jena, Germany.

出版信息

BMC Res Notes. 2011 Dec 6;4:525. doi: 10.1186/1756-0500-4-525.

Abstract

BACKGROUND

Brucellosis in livestock causes enormous losses for economies of developing countries and poses a severe health risk to consumers of dairy products. Little information is known especially on camel brucellosis and its impact on human health. For surveillance and control of the disease, sensitive and reliable detection methods are needed. Although serological tests are the mainstay of diagnosis in camel brucellosis, these tests have been directly transposed from cattle without adequate validation. To date, little information on application of real-time PCR for detection of Brucella in camel serum is available. Therefore, this study was performed to compare the diagnostic efficiency of different serological tests and real-time PCR in order to identify the most sensitive, rapid and simple combination of tests for detecting Brucella infection in camels.

FINDINGS

A total of 895 serum samples collected from apparently healthy Sudanese camels was investigated. Sudan is a well documented endemic region for brucellosis with cases in humans, ruminants, and camels. Rose Bengal Test (RBT), Complement Fixation Test (CFT), Slow Agglutination Test (SAT), Competitive Enzyme Linked Immunosorbant Assay (cELISA) and Fluorescence Polarization Assay (FPA) as well as real-time PCR were used. Our findings revealed that bcsp31 kDa real-time PCR detected Brucella DNA in 84.8% (759/895) of the examined samples, of which 15.5% (118/759) were serologically negative. Our results show no relevant difference in sensitivity between the different serological tests. FPA detected the highest number of positive cases (79.3%) followed by CFT (71.4%), RBT (70.7%), SAT (70.6%) and cELISA (68.8%). A combination of real-time PCR with one of the used serological tests identified brucellosis in more than 99% of the infected animals. 59.7% of the examined samples were positive in all serological tests and real-time PCR. A subpopulation of 6.8% of animals was positive in all serological tests but negative in real-time PCR assays. The high percentage of positive cases in this study does not necessarily reflect the seroprevalence of the disease in the country but might be caused by the fact that the camels were imported from brucellosis infected herds of Sudan, accidentally. Seroprevalence of brucellosis in camels should be examined in confirmatory studies to evaluate the importance of brucellosis in this animal species.

CONCLUSION

We suggest combining bcsp31 real-time PCR with either FPA, CFT, RBT or SAT to screen camels for brucellosis.

摘要

背景

家畜布鲁氏菌病给发展中国家的经济造成巨大损失,并对乳制品消费者构成严重健康风险。尤其是关于骆驼布鲁氏菌病及其对人类健康的影响,人们了解的信息很少。为了监测和控制该疾病,需要灵敏可靠的检测方法。虽然血清学检测是骆驼布鲁氏菌病诊断的主要手段,但这些检测方法直接照搬自牛的检测方法,未经过充分验证。迄今为止,关于实时荧光定量聚合酶链反应(PCR)在骆驼血清中检测布鲁氏菌的应用信息很少。因此,本研究旨在比较不同血清学检测和实时荧光定量PCR的诊断效率,以确定检测骆驼布鲁氏菌感染的最灵敏、快速和简便的检测组合。

研究结果

共调查了895份从表面健康的苏丹骆驼采集的血清样本。苏丹是一个布鲁氏菌病记录完备的流行地区,存在人类、反刍动物和骆驼感染病例。使用了玫瑰红试验(RBT)、补体结合试验(CFT)、迟缓凝集试验(SAT)、竞争酶联免疫吸附测定(cELISA)、荧光偏振测定(FPA)以及实时荧光定量PCR。我们的研究结果显示,bcsp31 kDa实时荧光定量PCR在84.8%(759/895)的检测样本中检测到布鲁氏菌DNA,其中15.5%(118/759)血清学检测为阴性。我们的结果表明不同血清学检测在灵敏度上没有显著差异。FPA检测到的阳性病例数最多(79.3%),其次是CFT(71.4%)、RBT(70.7%)、SAT(70.6%)和cELISA(68.8%)。实时荧光定量PCR与所使用的一种血清学检测方法联合使用,在超过99%的感染动物中检测出布鲁氏菌病。59.7%的检测样本在所有血清学检测和实时荧光定量PCR中均为阳性。6.8%的动物亚群在所有血清学检测中为阳性,但在实时荧光定量PCR检测中为阴性。本研究中高比例的阳性病例不一定反映该国该疾病的血清流行率,可能是因为这些骆驼是意外从苏丹感染布鲁氏菌病的畜群中引进的。骆驼布鲁氏菌病的血清流行率应在验证性研究中进行检测,以评估布鲁氏菌病在该动物物种中的重要性。

结论

我们建议将bcsp31实时荧光定量PCR与FPA、CFT、RBT或SAT联合使用,对骆驼进行布鲁氏菌病筛查。

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