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在源自波兰北部的欧洲鳗鲡(Anguilla anguilla (L.))中检测鳗疱疹病毒(AngHV-1)——对所选诊断方法适用性的评估

Detection of Herpesvirus anguillae (AngHV-1) in European eel Anguilla anguilla (L.) originating from northern Poland-assessment of suitability of selected diagnostic methods.

作者信息

Nguyen Tuan Thuc, Jin Yeonhwa, Kiełpińska Jolanta, Bergmann Sven M, Lenk Matthias, Panicz Remigiusz

机构信息

Department of Aquaculture, West Pomeranian University of Technology in Szczecin, Szczecin, Poland.

Faculty of Agriculture-Forestry-Fisheries, Vinh University, Vinh City, Nghe An province, Vietnam.

出版信息

J Fish Dis. 2017 Nov;40(11):1717-1723. doi: 10.1111/jfd.12689. Epub 2017 Aug 24.

Abstract

The Community Action Plan requests EU member states to implement measures that ensure the recovery of the severely depleted European eel stocks. One of the main threats is posed by Anguillid herpesvirus 1 (AngHV-1) leading to increased mortality in both wild and farmed eels. Following recommendations of the OIE to minimize the risk of obtaining false-negative results, the main aim of the study was to optimize diagnostic methods for AngHV-1 detection using conventional PCR, nested PCR and in situ hybridization assay. While 53.3% of the individual organ samples were tested positive for AngHV-1 by PCR, the additional virus analysis via nested PCR revealed that the actual prevalence was 93.3%. In the cell cultivation passages, a cytopathic effect was hardly found in the first two rounds. In the third passage onto cell cultures, a lytic CPE was detected. The identification and confirmation of the viruses obtained from cell cultures as well as directly from the organ tissues were proceeded by PCR, nested PCR and sequencing of the PCR products. While no positive signal was detectable in the first round by PCR using samples from the third cell culture passages, the nested PCR provided weak but visible positive signals.

摘要

《社区行动计划》要求欧盟成员国采取措施,确保严重枯竭的欧洲鳗鱼种群得以恢复。主要威胁之一是由鳗疱疹病毒1型(AngHV-1)造成的,它会导致野生和养殖鳗鱼的死亡率上升。按照世界动物卫生组织的建议,为尽量降低出现假阴性结果的风险,本研究的主要目的是优化使用常规PCR、巢式PCR和原位杂交检测法检测AngHV-1的诊断方法。通过PCR检测,53.3%的单个器官样本呈AngHV-1阳性,而通过巢式PCR进行的额外病毒分析显示实际感染率为93.3%。在细胞培养传代过程中,在前两轮几乎未发现细胞病变效应。在第三轮细胞培养传代时,检测到了溶细胞性细胞病变效应。通过PCR、巢式PCR以及对PCR产物进行测序,对从细胞培养物以及直接从器官组织中获得的病毒进行了鉴定和确认。使用来自第三轮细胞培养传代的样本进行PCR检测时,第一轮未检测到阳性信号,但巢式PCR提供了微弱但可见的阳性信号。

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