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基于系统进化分析和分子方法从黄鲈(Perca flavescens (Mitchell))中检测淋巴囊肿病病毒

Phylogenetic analysis and molecular methods for the detection of lymphocystis disease virus from yellow perch, Perca flavescens (Mitchell).

机构信息

Department of Biology, Canadian Rivers Institute, University of Prince Edward Island, Charlottetown, Prince Edward Island, Canada.

出版信息

J Fish Dis. 2012 Sep;35(9):661-70. doi: 10.1111/j.1365-2761.2012.01391.x. Epub 2012 Jul 15.

DOI:10.1111/j.1365-2761.2012.01391.x
PMID:22804739
Abstract

Lymphocystis disease is a prevalent, non-fatal disease that affects many teleost fish and is caused by the DNA virus lymphocystis disease virus (LCDV). Lymphocystis-like lesions have been observed in yellow perch, Perca flavescens (Mitchell), in lakes in northern Alberta, Canada. In an effort to confirm the identity of the virus causing these lesions, DNA was extracted from these lesions and PCR with genotype generic LCDV primers specific to the major capsid protein (MCP) gene was performed. A 1357-base pair nucleotide sequence corresponding to a peptide length of 452 amino acids of the MCP gene was sequenced, confirming the lesions as being lymphocystis disease lesions. Phylogenetic analysis of the generated amino acid sequence revealed the perch LCDV isolate to be a distinct and novel genotype. From the obtained sequence, a real-time PCR identification method was developed using fluorgenic LUX primers. The identification method was used to detect the presence/absence of LCDV in yellow perch from two lakes, one where lymphocystis disease was observed to occur and the other where the disease had not been observed. All samples of fin, spleen and liver tested negative for LCDV in the lake where lymphocystis disease had not been observed. The second lake had a 2.6% incidence of LCD, and virus was detected in tissue samples from all individuals tested regardless of whether they were expressing the disease or not. However, estimated viral copy number in spleen and liver of symptomatic perch was four orders of magnitude higher than that in asymptomatic perch.

摘要

淋巴囊肿病是一种普遍存在的非致命性疾病,影响许多硬骨鱼类,由 DNA 病毒淋巴囊肿病病毒 (LCDV)引起。在加拿大阿尔伯塔省北部的湖泊中,已观察到黄鲈(Perca flavescens (Mitchell))出现淋巴囊肿样病变。为了确认导致这些病变的病毒身份,从这些病变中提取 DNA,并使用针对主要衣壳蛋白 (MCP)基因的基因型通用 LCDV 引物进行 PCR。测序了与 MCP 基因的 452 个氨基酸长度相对应的 1357 个碱基对核苷酸序列,证实这些病变为淋巴囊肿病病变。生成的氨基酸序列的系统发育分析显示,鲈鱼 LCDV 分离株是一种独特的新型基因型。从获得的序列中,使用荧光 LUX 引物开发了实时 PCR 鉴定方法。该鉴定方法用于检测两个湖泊中黄鲈是否存在 LCDV,一个湖泊中观察到淋巴囊肿病的发生,另一个湖泊中未观察到该疾病。在未观察到淋巴囊肿病的湖中,所有鳍、脾和肝样本均未检测到 LCDV。第二个湖中淋巴囊肿病的发病率为 2.6%,无论是否表现出疾病,所有测试个体的组织样本中均检测到病毒。然而,在有症状的鲈鱼的脾和肝中的病毒拷贝数估计比无症状的鲈鱼高四个数量级。

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