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用于猫结核病的γ-干扰素酶联免疫吸附测定(ELISA)和酶联免疫斑点试验(ELISPOT)的适应性调整

Adaptation of IFN-gamma ELISA and ELISPOT tests for feline tuberculosis.

作者信息

Rhodes Shelley G, Gruffydd-Jones Tim, Gunn-Moore Danièlle, Jahans Keith

机构信息

TB Research Group, Veterinary Laboratories Agency, Surrey KT15 3NB, United Kingdom.

出版信息

Vet Immunol Immunopathol. 2008 Aug 15;124(3-4):379-84. doi: 10.1016/j.vetimm.2008.04.006. Epub 2008 Apr 16.

DOI:10.1016/j.vetimm.2008.04.006
PMID:18486239
Abstract

There are currently no reliable immunodiagnostic tests for feline tuberculosis. Infection of domestic cats in the UK is thought to occur via their contact with the relevant reservoir of infection, e.g. cattle and badgers for Mycobacterium bovis, and rodents for M. microti. In the African National Parks, where M. bovis infection of Bovidae is an increasing problem, transmission to big cats is occurring via their ingestion of infected carcasses. We have adapted feline ELISA and ELISPOT assays to potentially provide the first cell-based diagnostic test for the detection of tuberculosis in cats. We tested peripheral blood mononuclear cell antigen-specific IFN-gamma responses of 18 cats suspected of mycobacterial infection for which biopsy material was co-submitted to the Veterinary Laboratories Agency for mycobacterial culture and identification. Seventeen cats were tested by ELISA while seven cats were tested by ELISPOT (six cats were tested by both ELISA and ELISPOT). Six healthy control cats provided baseline data for these tests. Responses to bovine and avian tuberculins (PPDB and PPDA) and a protein cocktail of ESAT6 and CFP10 were measured, together with positive mitogen (PMA and calcium ionophore) and negative (medium) controls. Overall, both ELISPOT and ELISA tests were found to be suitable for generating rapid results (2 and 4 days, respectively), which provided good predictive information for M. bovis and M. microti infections, but were unable to reliably discern M. avium infection.

摘要

目前尚无用于猫结核病的可靠免疫诊断测试。英国的家猫感染被认为是通过与相关感染源接触而发生的,例如牛分枝杆菌的感染源是牛和獾,田鼠分枝杆菌的感染源是啮齿动物。在非洲国家公园,牛科动物感染牛分枝杆菌的问题日益严重,大型猫科动物通过摄食受感染的尸体而被传染。我们改进了猫酶联免疫吸附测定(ELISA)和酶联免疫斑点测定(ELISPOT),有可能为检测猫结核病提供首个基于细胞的诊断测试。我们检测了18只疑似分枝杆菌感染猫的外周血单核细胞抗原特异性γ干扰素反应,这些猫的活检材料同时提交给兽医实验室机构进行分枝杆菌培养和鉴定。17只猫通过ELISA检测,7只猫通过ELISPOT检测(6只猫同时通过ELISA和ELISPOT检测)。6只健康对照猫为这些测试提供了基线数据。测量了对牛结核菌素和禽结核菌素(PPDB和PPDA)以及早期分泌性抗原靶6(ESAT6)和培养滤液蛋白10(CFP10)的蛋白混合物的反应,同时设置了阳性促细胞分裂剂(佛波酯和离子霉素)和阴性(培养基)对照。总体而言,发现ELISPOT和ELISA测试都适合快速得出结果(分别为2天和4天),这为牛分枝杆菌和田鼠分枝杆菌感染提供了良好的预测信息,但无法可靠地辨别鸟分枝杆菌感染。

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