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应用原位检测技术确定养殖金头鲷(Sparus aurata L.)中淋巴囊肿病病毒感染的全身状况

Application of in situ detection techniques to determine the systemic condition of lymphocystis disease virus infection in cultured gilt-head seabream, Sparus aurata L.

作者信息

Cano I, Ferro P, Alonso M C, Sarasquete C, Garcia-Rosado E, Borrego J J, Castro D

机构信息

Departamento de Microbiología, Facultad de Ciencias, Universidad de Málaga, Málaga, Spain.

出版信息

J Fish Dis. 2009 Feb;32(2):143-50. doi: 10.1111/j.1365-2761.2008.00970.x. Epub 2008 Sep 18.

Abstract

Immunohistochemistry (IHC) and in situ hybridization (ISH) techniques have been used for the detection of lymphocystis disease virus (LCDV) in formalin-fixed, paraffin-embedded tissues from gilt-head seabream, Sparus aurata L. Diseased and recovered fish from the same population were analysed. IHC was performed with a polyclonal antibody against a 60-kDa viral protein. A specific digoxigenin-labelled probe, obtained by PCR amplification of a 270-bp fragment of the gene coding the LCDV major capsid protein, was used for ISH. LCDV was detected in skin dermis and gill lamellae, as well as in several internal organs such as the intestine, liver, spleen and kidney using both techniques. Fibroblasts, hepatocytes and macrophages seem to be target cells for virus replication. The presence of lymphocystis cells in the dermis of the skin and caudal fin, and necrotic changes in the epithelium of proximal renal tubules were the only histological alterations observed in fish showing signs of the disease.

摘要

免疫组织化学(IHC)和原位杂交(ISH)技术已用于检测来自金头鲷(Sparus aurata L.)的福尔马林固定、石蜡包埋组织中的淋巴囊肿病病毒(LCDV)。对来自同一群体的患病和康复鱼类进行了分析。使用针对一种60 kDa病毒蛋白的多克隆抗体进行免疫组织化学检测。通过对编码LCDV主要衣壳蛋白的基因的270 bp片段进行PCR扩增获得的一种地高辛标记的特异性探针用于原位杂交。使用这两种技术在皮肤真皮、鳃小片以及肠道、肝脏、脾脏和肾脏等几个内部器官中检测到了LCDV。成纤维细胞、肝细胞和巨噬细胞似乎是病毒复制的靶细胞。在出现疾病症状的鱼类中观察到的唯一组织学改变是皮肤和尾鳍真皮中存在淋巴囊肿细胞以及近端肾小管上皮的坏死变化。

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