Shimatsu Taichi, Shinozaki Harumi, Kimitsuki Kazunori, Shiwa Nozomi, Manalo Daria L, Perez Rodolfo C, Dilig Joselito E, Yamada Kentaro, Boonsriroj Hassadin, Inoue Satoshi, Park Chun-Ho
Department of Veterinary Pathology, School of Veterinary Medicine, Kitasato University, 23-35-1, Higashi, Towada, Aomori 034-8628, Japan.
Veterinary Research Department, Research Institute for Tropical Medicine, Department of Health, 9002 Research Drive, Filinvest Corporate City, Alabang, Muntinlupa City 1781, Philippines.
J Virol Methods. 2016 Nov;237:40-46. doi: 10.1016/j.jviromet.2016.08.021. Epub 2016 Aug 30.
The direct fluorescent antibody test (dFAT) on fresh brain tissues is the gold standard for rabies virus antigen detection in dogs. However, this method is laborious and holds a high risk of virus exposure for the experimenter. Skin biopsies are useful for the diagnosis of humans and animals. In mammals, the tactile hair, known as the follicle-sinus complex (FSC), is a specialized touch organ that is abundant in the muzzle skin. Each tactile hair is equipped with more than 2,000 sensory nerve endings. Therefore, this organ is expected to serve as an alternative postmortem diagnostic material. However, the target cells and localization of rabies virus antigen in the FSCs remain to be defined. In the present study, muzzle skins were obtained from 60 rabid dogs diagnosed with rabies by dFAT at the Research Institute of Tropical Medicine in the Philippines. In all dogs, virus antigen was clearly detected in a part of the outer root sheath at the level of the ring sinus of the FSCs, and the majority of cells were positive for the Merkel cell (MC) markers cytokeratin 20 and CAM5.2. Our results suggest that MCs in the FSCs of the muzzle skin are a target for virus replication and could serve as a useful alternative specimen source for diagnosis of rabies.
对新鲜脑组织进行直接荧光抗体试验(dFAT)是检测犬类狂犬病病毒抗原的金标准。然而,该方法操作繁琐,且实验人员面临较高的病毒暴露风险。皮肤活检对人和动物的诊断都很有用。在哺乳动物中,触觉毛发,即毛囊-窦复合体(FSC),是一种特殊的触觉器官,在口鼻部皮肤中大量存在。每根触觉毛发都配备有2000多个感觉神经末梢。因此,这个器官有望作为一种替代性的死后诊断材料。然而,狂犬病病毒抗原在FSCs中的靶细胞和定位仍有待确定。在本研究中,从菲律宾热带医学研究所通过dFAT诊断为狂犬病的60只狂犬的口鼻部皮肤中获取样本。在所有犬只中,在FSCs环形窦水平的外根鞘的一部分中均清晰检测到病毒抗原,并且大多数细胞对默克尔细胞(MC)标记物细胞角蛋白20和CAM5.2呈阳性。我们的结果表明,口鼻部皮肤FSCs中的MCs是病毒复制的靶标,并且可以作为狂犬病诊断的有用替代标本来源。