A new homogeneous assay for high throughput serological diagnosis of brucellosis in ruminants
A new homogeneous assay for high throughput serological diagnosis of brucellosis in ruminants
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DOI：
10.1016/j.jim.2008.05.004
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发表时间：
2008-08-20
影响因子：
2.2
通讯作者：
Stack, Judy A.
中科院分区：
文献类型：
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作者：
McGiven, John A.;Sawyer, Jason;Stack, Judy A.
布鲁氏菌病的控制和根除是非常理想的，但由于需要高通量血清学检测，因此需要大量资源。本研究的目的是通过开发一种新的检测方法来满足参与该过程的高通量筛选实验室的需求。将用于反刍动物布鲁氏菌病血清学诊断的现有cELISA转换为AlphaLISA均相邻近检测。该测定不需要分离步骤，并且可以以低体积微量滴定形式进行。在一组来自感染和未感染动物的牛、绵羊和山羊血清中验证了布鲁氏菌AlphaLISA。所获得的诊断灵敏度（>96%）和特异性（>98%）与来自对相同样品进行的cELISA、iELISA和FPA的诊断灵敏度（> 96%）和特异性（>98%）相比良好。AlphaLISA符合OIEELISA标准品定义的ELISA检测标准，分析灵敏度与母体cELISA相似。该方法还用于一小组来自实验感染小肠结肠炎耶尔森氏菌O：9的牛的血清样品。获得了一些假阳性反应，FPA、iELISA、cELISA、CFT和SAT的结果也是如此。尽管如此，AlphaLISA的方法学优势意味着该检测非常适合高通量血清学诊断。本报告是首次描述使用AlphaLISA检测病原体特异性抗体。此外，cELISA转换到该平台的相对容易性表明，该技术已准备好满足许多其他疾病诊断的高通量测试要求。(C)2008 Elsevier B.V.保留所有权利。
The control and eradication of brucellosis is highly desirable but heavily resource intensive as high throughput serological testing is required. The aim of this study was to meet the needs of high throughput screening laboratories involved in this process through the development of a new assay. An existing cELISA used for the serodiagnosis of brucellosis in ruminants was converted to an AlphaLISA homogenous proximity based assay. This assay requires no separation steps and can be performed in low volume microtitre format. The Brucella AlphaLISA was validated on a panel of bovine, ovine and caprine sera from infected and uninfected animals. The diagnostic sensitivities (>96%) and specificities (>98%) obtained compared well to those from cELISA, iELISA and FPA performed on the same samples. The AlphaLISA met the testing criteria set for ELISAs as defined by the OIEELISA standards and had an analytical sensitivity similar to that of the parent cELISA. The method was also used on a small panel of serum samples from cattle that were experimentally infected with Yersinia enterocolitica O:9. Some false positive reactions were obtained as was also the case with results from FPA, iELISA, cELISA, CFT and SAT. Despite this, the methodological advantages of the AlphaLISA mean that this assay is well suited to high throughput serodiagnosis. This report is the first description of the use of AlphaLISA to detect pathogen specific antibodies. Furthermore, the relative ease with which the cELISA was converted to this platform indicates that this technology is ready to meet the high throughput testing requirements for the diagnosis of many other diseases. (C) 2008 Elsevier B.V. All rights reserved.