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与马昆虫叮咬超敏反应诊断相关的骚扰库蠓提取物。

Culicoides obsoletus extract relevant for diagnostics of insect bite hypersensitivity in horses.

作者信息

van der Meide Nathalie M A, Meulenbroeks Chantal, van Altena Christine, Schurink Anouk, Ducro Bart J, Wagner Bettina, Leibold Wolfgang, Rohwer Jens, Jacobs Frans, van Oldruitenborgh-Oosterbaan Marianne M Sloet, Savelkoul Huub F J, Tijhaar Edwin

机构信息

Cell Biology and Immunology Group, Wageningen University, Wageningen, The Netherlands.

出版信息

Vet Immunol Immunopathol. 2012 Oct 15;149(3-4):245-54. doi: 10.1016/j.vetimm.2012.07.007. Epub 2012 Jul 23.

DOI:10.1016/j.vetimm.2012.07.007
PMID:22906994
Abstract

Insect bite hypersensitivity (IBH) is an allergic dermatitis in horses caused by the bites of Culicoides species. The aim of the present study was to evaluate the applicability of whole body extracts of C. obsoletus (the main species found feeding on horses in the Netherlands), C. nubeculosus (rarely found in The Netherlands) and C. sonorensis (typical for North America) for diagnosis of IBH in horses in The Netherlands. Blood and serum samples of 10 clinically confirmed IBH affected and 10 healthy control horses were used to evaluate the IgE titers (ELISA) against the Culicoides whole body extracts of the three Culicoides species. Basophil degranulation was assessed by histamine release test (HRT) after stimulation with these extracts at 5, 0.5 and 0.05 μg/ml. IBH affected horses had significantly higher IgE titers against C. obsoletus than against C. nubeculosus and C. sonorensis. Furthermore, C. obsoletus induced significantly higher histamine release in whole blood of IBH affected horses compared to the other extracts at 0.5 μg/ml. Western blot data revealed IgE binding to many proteins in C. obsoletus extract. This interaction was absent or weak in C. nubeculosus and C. sonorensis extracts for IBH affected horses. Results on individual level indicate that the HRT is more sensitive than ELISA in diagnosing IBH. However, ELISA is more practical as a routine test, therefore the ELISA was further evaluated using C. obsoletus extract on 103 IBH affected and 100 healthy horses, which resulted in a test sensitivity and specificity of 93.2% and 90.0%, respectively. The IgE ELISA readings enabled the analysis of the predicted probability of being IBH affected. From an optical density 450nm value of 0.33 onwards, the probability of IBH affected was more than 0.9. The results presented in this paper show that the use of native Culicoides spp. that feed on horse, is important for improved diagnosis and that the described ELISA based on C. obsoletus can be used routinely to diagnose IBH in countries where this species is the main Culicoides feeding on horses.

摘要

昆虫叮咬超敏反应(IBH)是一种由库蠓属昆虫叮咬引起的马过敏性皮炎。本研究的目的是评估荷兰常见的骚扰库蠓、较少见的云雾库蠓以及北美洲典型的索诺拉库蠓的全虫提取物在荷兰马匹IBH诊断中的适用性。使用10匹临床确诊为IBH的患病马和10匹健康对照马的血液及血清样本,评估针对这三种库蠓全虫提取物的IgE滴度(酶联免疫吸附测定法)。在用浓度为5、0.5和0.05μg/ml的这些提取物刺激后,通过组胺释放试验(HRT)评估嗜碱性粒细胞脱颗粒情况。患IBH的马针对骚扰库蠓的IgE滴度显著高于针对云雾库蠓和索诺拉库蠓的IgE滴度。此外,在浓度为0.5μg/ml时,与其他提取物相比,骚扰库蠓在患IBH马的全血中诱导的组胺释放显著更高。蛋白质印迹数据显示IgE与骚扰库蠓提取物中的许多蛋白质结合。对于患IBH的马,这种相互作用在云雾库蠓和索诺拉库蠓提取物中不存在或较弱。个体水平的结果表明,在诊断IBH方面,HRT比酶联免疫吸附测定法更敏感。然而,酶联免疫吸附测定法作为常规检测更实用,因此使用骚扰库蠓提取物对103匹患IBH的马和100匹健康马进一步评估酶联免疫吸附测定法,结果显示检测灵敏度和特异性分别为93.2%和90.0%。IgE酶联免疫吸附测定读数能够分析患IBH的预测概率。从450nm光密度值0.33起,患IBH 的概率超过0.9。本文给出的结果表明,使用以马为食的本地库蠓物种对于改善诊断很重要,并且基于骚扰库蠓所描述的酶联免疫吸附测定法可在该物种为以马为食的主要库蠓的国家常规用于诊断IBH。

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