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在红斑赤虾(樱虾科)中鉴定一种虹彩病毒及其原位杂交和PCR检测方法的建立。

Identification of an iridovirus in Acetes erythraeus (Sergestidae) and the development of in situ hybridization and PCR method for its detection.

作者信息

Tang Kathy F J, Redman Rita M, Pantoja Carlos R, Groumellec Marc Le, Duraisamy Panchayuthapani, Lightner Donald V

机构信息

Department of Veterinary Science and Microbiology, University of Arizona, Tucson, AZ 85721, USA.

出版信息

J Invertebr Pathol. 2007 Nov;96(3):255-60. doi: 10.1016/j.jip.2007.05.006. Epub 2007 May 18.

Abstract

An iridovirus (tentatively named SIV, sergestid iridovirus) that causes high mortality in the sergestid shrimp, Acetes erythraeus, was found in Madagascar in 2004. Severely affected shrimp exhibit a blue-green opalescence. Histological examination revealed massive cytoplasmic inclusions in the cuticular epithelial cells, connective tissues, ovary and testes. The electron microscopic examination showed paracrystalline arrays of virions at a size of 140nm, suggesting infection with an iridovirus. A pair of PCR primers were selected from the conserved region of the major capsid protein (MCP)-coding sequence among insect iridoviruses and used to amplify a 1.0kb fragment from the infected A. erythraeus. This fragment was cloned, sequenced and found to be highly similar (upto 80% similarity in translated amino acids with an E value of 1e-124) to the MCP of invertebrate iridoviruses. This clone was then labeled with digoxigenin-11-dUTP and hybridized to tissue sections of infected A. erythraeus, which reacted positively to the probe. The reacting tissues included epithelial cells, connective tissues, and the germinal cells; the same cells as those with inclusions. A PCR method was also developed from the MCP coding sequence for detecting SIV.

摘要

2004年在马达加斯加发现了一种虹彩病毒(暂命名为SIV,十足目虹彩病毒),该病毒在红斑赤虾中可导致高死亡率。受严重影响的虾呈现蓝绿色的珠光。组织学检查显示,角质上皮细胞、结缔组织、卵巢和睾丸中存在大量细胞质内含物。电子显微镜检查显示,病毒粒子呈140纳米大小的准晶体阵列,表明感染了虹彩病毒。从昆虫虹彩病毒主要衣壳蛋白(MCP)编码序列的保守区域中选择了一对PCR引物,用于从受感染的红斑赤虾中扩增出一个1.0kb的片段。该片段被克隆、测序,发现与无脊椎动物虹彩病毒的MCP高度相似(翻译后的氨基酸相似度高达80%,E值为1e-124)。然后用洋地黄毒苷-11-dUTP标记该克隆,并与受感染的红斑赤虾组织切片杂交,组织切片与探针呈阳性反应。发生反应的组织包括上皮细胞、结缔组织和生殖细胞;与有内含物的细胞相同。还从MCP编码序列开发了一种用于检测SIV的PCR方法。

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