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狮特异性γ干扰素检测方法的开发。

Development of a lion-specific interferon-gamma assay.

作者信息

Maas M, van Kooten P J S, Schreuder J, Morar D, Tijhaar E, Michel A L, Rutten V P M G

机构信息

Division of Epidemiology, Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 7, 3584 CL Utrecht, The Netherlands.

出版信息

Vet Immunol Immunopathol. 2012 Oct 15;149(3-4):292-7. doi: 10.1016/j.vetimm.2012.07.014. Epub 2012 Aug 4.

Abstract

The ongoing spread of bovine tuberculosis (BTB) in African free-ranging lion populations, for example in the Kruger National Park, raises the need for diagnostic assays for BTB in lions. These, in addition, would be highly relevant for zoological gardens worldwide that want to determine the BTB status of their lions, e.g. for translocations. The present study concerns the development of a lion-specific IFN-γ assay, following the production and characterization of monoclonal antibodies specific for lion interferon-gamma (IFN-γ). Recombinant lion IFN-γ (rLIFN-γ) was produced in mammalian cells and used to immunize mice to establish hybridoma cell lines producing monoclonal antibodies. These were used to develop a sensitive, lion IFN-γ-specific capture ELISA, able to detect rLIFN-γ to the level of 160 pg/ml. Recognition of native lion IFN-γ was shown in an initial assessment of supernatants of mitogen stimulated whole blood cultures of 11 known BTB-negative lions. In conclusion, the capture ELISA shows potential as a diagnostic assay for bovine tuberculosis in lions. Preliminary results also indicate the possible use of the test for other (feline) species.

摘要

牛结核病(BTB)在非洲自由放养的狮子种群中持续传播,例如在克鲁格国家公园,这就需要开发针对狮子的BTB诊断检测方法。此外,对于全球范围内想要确定其狮子的BTB状况(例如用于转移)的动物园来说,这些检测方法也具有高度相关性。本研究涉及一种针对狮子的干扰素-γ(IFN-γ)检测方法的开发,此前已制备并鉴定了针对狮子干扰素-γ(IFN-γ)的单克隆抗体。重组狮子IFN-γ(rLIFN-γ)在哺乳动物细胞中产生,并用于免疫小鼠以建立产生单克隆抗体的杂交瘤细胞系。这些单克隆抗体被用于开发一种灵敏的、针对狮子IFN-γ的捕获ELISA,能够检测到低至160 pg/ml水平的rLIFN-γ。在对11只已知BTB阴性狮子的有丝分裂原刺激全血培养上清液的初步评估中,显示了对天然狮子IFN-γ的识别。总之,捕获ELISA显示出作为狮子牛结核病诊断检测方法有潜力。初步结果还表明该检测方法可能适用于其他(猫科)物种。

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