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用于检测法国和美国企鹅中抗体的商业化 Western blot 试剂盒的适应性改造。

ADAPTATION OF A COMMERCIALLY AVAILABLE WESTERN BLOT KIT FOR THE DETECTION OF ANTIBODY TO IN PENGUINS IN FRANCE AND THE UNITED STATES.

机构信息

ZooParc de Beauval, 41110 Saint-Aignan, France,

LDBIO Diagnostics, 69009 Lyon, France.

出版信息

J Zoo Wildl Med. 2024 Sep;55(3):595-601. doi: 10.1638/2024-0008.

Abstract

Antemortem serodiagnosis of aspergillosis remains challenging in Sphenisciformes. Protein electrophoresis, serology (antibody, antigen) by ELISA, and gliotoxin detection provide variable diagnostic value. In the present study, a commercially available Western blot (WB) validated for use in humans and dolphins was adapted for use with penguin samples. Using the same method and reagents, samples were analyzed from multiple institutions in the United States and one facility in France. This was inclusive of normal juvenile African penguins (, n = 10) and various species of penguins in the United States with confirmed infection (n = 9) as well as 52 samples from Humboldt penguins () in France. Cumulative WB scores (based on reactivity to different antigens) were found to be significantly higher in the group of penguins with confirmed infection (p < 0.0001). Significant differences were also observed between the clinically normal penguins in the two populations, with higher scores in the United States (median score 1.0, 95%CI [0-5], min 0, max 11) compared to France (median score 0,95%CI [0-0], min 0, max 5). The utilization of the WB as a diagnostic tool is inconclusive due to the use of samples from varying institutions, environmental background, age, and stages of infection. However, this tool may provide an overview of antigen reactivity in penguins infected with to help design a more robust serology assay and further understand the humoral immune response during infection.

摘要

在企鹅目中,生前曲霉病的血清学诊断仍然具有挑战性。蛋白质电泳、酶联免疫吸附试验(ELISA)的血清学(抗体、抗原)和Gliotoxin 检测提供了不同的诊断价值。在本研究中,我们对一种已商业化并经人类和海豚验证的 Western blot(WB)进行了适应性修改,以用于企鹅样本。使用相同的方法和试剂,我们分析了来自美国多个机构和法国一个设施的样本。这包括正常的幼年非洲企鹅(n=10)和美国各种已确认感染的企鹅物种(n=9),以及法国的 52 只洪堡企鹅(n=52)样本。发现具有确认感染的企鹅组的累积 WB 评分(基于对不同抗原的反应性)显著更高(p<0.0001)。在两个种群的临床正常企鹅之间也观察到了显著差异,美国的评分更高(中位数评分 1.0,95%CI [0-5],最小值 0,最大值 11),而法国的评分较低(中位数评分 0,95%CI [0-0],最小值 0,最大值 5)。由于使用了来自不同机构、环境背景、年龄和感染阶段的样本,WB 作为诊断工具的应用尚无定论。然而,该工具可能提供了感染 后企鹅的抗原反应概述,有助于设计更强大的血清学检测方法,并进一步了解感染期间的体液免疫反应。

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