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用于检测密苏里河鲟人工养殖群体和野生群体中虹彩病毒感染的聚合酶链式反应检测方法的开发。

Development of PCR assays to detect iridovirus infections among captive and wild populations of Missouri River sturgeon.

作者信息

Kurobe T, Kwak K T, MacConnell E, McDowell T S, Mardones F O, Hedrick R P

机构信息

Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, California 95616, USA.

出版信息

Dis Aquat Organ. 2010 Dec 7;93(1):31-42. doi: 10.3354/dao02284.

DOI:10.3354/dao02284
PMID:21290894
Abstract

The Missouri River sturgeon iridovirus (MRSIV) is an important factor contributing to losses during the hatchery rearing of juvenile pallid Scaphirhynchus albus and shovelnose S. platorynchus sturgeon. As the virus has not been isolated in cell culture, current detection procedures rely upon a combination of light and electron microscopy. Detection of characteristic virus-infected cells in the integument, usually of the fins, in hematoxylin and eosin (H&E)-stained tissue sections provides a presumptive finding. Confirmation requires observation by electron microscopy of characteristic doubly enveloped hexagonal virions of the appropriate size in the host cell cytoplasm. To improve these diagnostic procedures, a conventional polymerase chain reduction (PCR) assay was developed as a sensitive and specific method for detection of MRSIV DNA as found in numerous tissues of both naturally and experimentally infected pallid and shovelnose sturgeon. Sequences of amplicons obtained from testing of wild-caught shovelnose sturgeon and juvenile pallid sturgeon during hatchery outbreaks were identical, suggesting that the viruses found in both sturgeon are similar or closely related. In addition, a TaqMan PCR was developed that allowed estimates of the concentrations of MRSIV DNA present in the tissues of pallid and shovelnose sturgeon during acute and persistent infection. These new PCR assays are improved methods to detect MRSIV, but equally importantly, they provide insights into to the biology of the agent for more effective management of viral diseases in captive and wild Missouri River sturgeon populations.

摘要

密苏里河鲟鱼虹彩病毒(MRSIV)是导致浅色铲鲟(Scaphirhynchus albus)和铲鼻鲟(S. platorynchus)幼鱼在孵化场养殖过程中出现损失的一个重要因素。由于该病毒尚未在细胞培养中分离出来,目前的检测程序依赖于光学显微镜和电子显微镜相结合的方法。在苏木精和伊红(H&E)染色的组织切片中,检测到通常在鳍部的体表有特征性病毒感染细胞,可提供一个初步诊断结果。确诊需要通过电子显微镜观察宿主细胞质中具有适当大小的特征性双包膜六边形病毒粒子。为了改进这些诊断程序,开发了一种常规聚合酶链反应(PCR)检测方法,作为一种灵敏且特异的方法,用于检测在自然感染和实验感染的浅色铲鲟和铲鼻鲟的多种组织中发现的MRSIV DNA。在孵化场疫情期间,对野生捕获的铲鼻鲟和浅色铲鲟幼鱼进行检测获得的扩增子序列相同,这表明在两种鲟鱼中发现的病毒相似或密切相关。此外,还开发了一种TaqMan PCR方法,可对急性和持续性感染期间浅色铲鲟和铲鼻鲟组织中存在的MRSIV DNA浓度进行估计。这些新的PCR检测方法是检测MRSIV的改进方法,但同样重要的是,它们为了解该病原体的生物学特性提供了见解,有助于更有效地管理圈养和野生密苏里河鲟鱼种群中的病毒性疾病。

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