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石蜡包埋组织中猪霍乱病毒糖蛋白55的免疫组织化学检测

Immunohistochemical detection of hog cholera viral glycoprotein 55 in paraffin-embedded tissues.

作者信息

de las Mulas J M, Ruiz-Villamor E, Donoso S, Quezada M, Lecocq C, Sierra M A

机构信息

Department of Comparative Pathology, Veterinary Faculty, University of Córdoba, Spain.

出版信息

J Vet Diagn Invest. 1997 Jan;9(1):10-6. doi: 10.1177/104063879700900103.

Abstract

Formalin-fixed, paraffin embedded tissues obtained from 40 pigs inoculated with a field isolate of hog cholera virus were examined for the presence of Gp55, a major structural protein of the virus envelope, using a monoclonal antibody-based immunohistochemical test with the avidin-biotin-peroxidase complex method. Immunoreactivity was detected in hog cholera virus-infected tissues but not in control pigs tissues, African swine fever virus-infected tissues, or bovine viral diarrhea virus-infected porcine or bovine tissues. The first positive reactions were seen in lymphatic tissues, digestive tract and skin on postinoculation day (pid) 4, respiratory and urinary tissues on pid 5, nervous tissues on pid 6, and endocrine tissues on pid 7. These staining reactions persisted until the last observation on pid 18. Hog cholera virus antigen was not detected in heart tissue at any time. The highest levels of antigen detection were found in tonsils, spleen, and pancreas, although the esophageal mucosa and skin epithelial cells were also intensely and widely stained. The cellular staining pattern of Gp55 had a ubiquitous distribution. It was found in epithelial cells, macrophages and circulating monocytes, endothelial cells, lymphoid cells, and glial cells. The results showed a high specificity and high sensitivity for detecting hog cholera Gp55 in formalin-fixed, paraffin embedded tissue samples. This method allows precise association of Gp55 with specific cells, tissues, and histologic lesions, making the technique suitable for use in routine diagnosis of hog cholera.

摘要

使用基于单克隆抗体的免疫组织化学检测方法和抗生物素蛋白-生物素-过氧化物酶复合物法,对从40头接种猪霍乱病毒野毒株的猪身上获取的福尔马林固定、石蜡包埋组织进行检测,以确定病毒包膜主要结构蛋白Gp55的存在情况。在猪霍乱病毒感染的组织中检测到免疫反应性,但在对照猪组织、非洲猪瘟病毒感染的组织或牛病毒性腹泻病毒感染的猪或牛组织中未检测到。首次阳性反应出现在接种后第4天(pid)的淋巴组织、消化道和皮肤,第5天的呼吸道和泌尿组织,第6天的神经组织,以及第7天的内分泌组织。这些染色反应一直持续到接种后第18天的最后一次观察。在任何时候心脏组织中均未检测到猪霍乱病毒抗原。尽管食管黏膜和皮肤上皮细胞也被强烈且广泛地染色,但抗原检测水平最高的是扁桃体、脾脏和胰腺。Gp55的细胞染色模式分布广泛。它存在于上皮细胞、巨噬细胞和循环单核细胞、内皮细胞、淋巴细胞和神经胶质细胞中。结果表明,该方法在福尔马林固定、石蜡包埋组织样本中检测猪霍乱Gp55具有高特异性和高灵敏度。该方法能够使Gp55与特定细胞、组织和组织学病变精确关联,使得该技术适用于猪霍乱的常规诊断。

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