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从存档的 Davidson 固定石蜡包埋虾组织中检测到桃拉综合征病毒并进行系统进化分析。

Detection and Phylogenetic Analyses of Taura Syndrome Virus from Archived Davidson's-Fixed Paraffin-Embedded Shrimp Tissue.

机构信息

Aquaculture Pathology Laboratory, School of Animal and Comparative Biomedical Sciences, University of Arizona, Tucson, AZ 85721, USA.

出版信息

Viruses. 2020 Sep 16;12(9):1030. doi: 10.3390/v12091030.


DOI:10.3390/v12091030
PMID:32948008
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7551232/
Abstract

Taura syndrome is a World Organization for Animal Health (OIE)-listed disease of marine shrimp that is caused by Taura syndrome virus (TSV), a single-stranded RNA virus. Here we demonstrate the utility of using 15-year-old archived Davidson's-fixed paraffin-embedded (DFPE) shrimp tissues for TSV detection and phylogenetic analyses. Total RNA was isolated from known TSV-infected DFPE tissues using three commercially available kits and the purity and ability to detect TSV in the isolated RNA were compared. TSV was successfully detected through RT-qPCR in all the tested samples. Among the TSV-specific primers screened through RT-PCR, primer pair TSV-20 for the RNA-dependent RNA polymerase (RdRp), primers TSV-15 and TSV-16 for the capsid protein gene VP2 and primers TSV-5 for the capsid protein gene VP1 amplified the highest number of samples. To assess the phylogenetic relation among different TSV isolates, the VP1 gene was amplified and sequenced in overlapping segments. Concatenated sequences from smaller fragments were taken for phylogenetic analyses. The results showed that the TSV isolates from this study generally clustered with homologous isolates from the corresponding geographical regions indicating RNA derived from DFPE tissues can be used for pathogen detection and retrospective analyses. The ability to perform genomic characterization from archived tissue will expedite pathogen discovery, development of diagnostic tools and prevent disease spread in shrimp and potentially other aquaculture species worldwide.

摘要

桃拉综合征是世界动物卫生组织(OIE)列出的一种海洋虾类疾病,由桃拉综合征病毒(TSV)引起,这是一种单链 RNA 病毒。在这里,我们展示了使用 15 年前存档的 Davidson 固定石蜡包埋(DFPE)虾组织进行 TSV 检测和系统发育分析的实用性。使用三种市售试剂盒从已知 TSV 感染的 DFPE 组织中分离总 RNA,并比较了分离 RNA 的纯度和检测 TSV 的能力。通过 RT-qPCR 在所有测试样本中均成功检测到 TSV。在通过 RT-PCR 筛选的 TSV 特异性引物中,用于 RNA 依赖性 RNA 聚合酶(RdRp)的引物对 TSV-20、用于衣壳蛋白基因 VP2 的引物 TSV-15 和 TSV-16 以及用于衣壳蛋白基因 VP1 的引物 TSV-5 扩增了最多数量的样本。为了评估不同 TSV 分离株之间的系统发育关系,通过 RT-PCR 扩增并测序了 VP1 基因的重叠片段。来自较小片段的拼接序列用于系统发育分析。结果表明,本研究中的 TSV 分离株通常与相应地理区域的同源分离株聚类,表明源自 DFPE 组织的 RNA 可用于病原体检测和回顾性分析。从存档组织中进行基因组特征分析的能力将加速病原体的发现、诊断工具的开发,并防止虾类和全球潜在其他水产养殖物种中的疾病传播。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0953/7551232/3c1b8f75fb79/viruses-12-01030-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0953/7551232/60f7aa6284e8/viruses-12-01030-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0953/7551232/b418a79d836b/viruses-12-01030-g002a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0953/7551232/6c3cd42928f8/viruses-12-01030-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0953/7551232/b345921dddff/viruses-12-01030-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0953/7551232/0019e3452519/viruses-12-01030-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0953/7551232/3c1b8f75fb79/viruses-12-01030-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0953/7551232/60f7aa6284e8/viruses-12-01030-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0953/7551232/b418a79d836b/viruses-12-01030-g002a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0953/7551232/6c3cd42928f8/viruses-12-01030-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0953/7551232/b345921dddff/viruses-12-01030-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0953/7551232/0019e3452519/viruses-12-01030-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0953/7551232/3c1b8f75fb79/viruses-12-01030-g006.jpg

相似文献

[1]
Detection and Phylogenetic Analyses of Taura Syndrome Virus from Archived Davidson's-Fixed Paraffin-Embedded Shrimp Tissue.

Viruses. 2020-9-16

[2]
Complete genome reconstruction and genetic analysis of Taura syndrome virus of shrimp from archival Davidson's-fixed paraffin embedded tissue.

Virology. 2021-1-15

[3]
Reverse transcription polymerase chain reaction (RT-PCR) used for the detection of Taura syndrome virus (TSV) in experimentally infected shrimp.

Dis Aquat Organ. 1998-10-8

[4]
Characterization of a new strain of Taura syndrome virus (TSV) from Colombian shrimp farms and the implication in the selection of TSV resistant lines.

J Invertebr Pathol. 2012-9-25

[5]
Detection of Taura syndrome virus (TSV) strain differences using selected diagnostic methods: diagnostic implications in penaeid shrimp.

Dis Aquat Organ. 2002-11-7

[6]
A new RNA-friendly fixative for the preservation of penaeid shrimp samples for virological detection using cDNA genomic probes.

J Virol Methods. 1997-7

[7]
Simultaneous detection of white spot syndrome virus (WSSV) and Taura syndrome virus (TSV) by multiplex reverse transcription-polymerase chain reaction (RT-PCR) in pacific white shrimp Penaeus vannamei.

Dis Aquat Organ. 2002-6-21

[8]
Taura syndrome virus from Penaeus vannamei shrimp cultured in Korea.

Dis Aquat Organ. 2006-6-12

[9]
Quantitative assay for measuring the Taura syndrome virus and yellow head virus load in shrimp by real-time RT-PCR using SYBR Green chemistry.

J Virol Methods. 2002-6

[10]
Comparison of four Taura syndrome virus (TSV) isolates in oral challenge studies with Litopenaeus vannamei unselected or selected for resistance to TSV.

Dis Aquat Organ. 2006-7-11

引用本文的文献

[1]
Microbes and pathogens associated with shrimps - implications and review of possible control strategies.

Front Mar Sci. 2024

[2]
Viral Shrimp Diseases Listed by the OIE: A Review.

Viruses. 2022-3-12

本文引用的文献

[1]
Genome reconstruction of white spot syndrome virus (WSSV) from archival Davidson's-fixed paraffin embedded shrimp (Penaeus vannamei) tissue.

Sci Rep. 2020-8-10

[2]
FFPE breast tumour blocks provide reliable sources of both germline and malignant DNA for investigation of genetic determinants of individual tumour responses to treatment.

Breast Cancer Res Treat. 2018-4-26

[3]
ICTV Virus Taxonomy Profile: Dicistroviridae.

J Gen Virol. 2017-3

[4]
Virus characterization and discovery in formalin-fixed paraffin-embedded tissues.

J Virol Methods. 2015-3

[5]
Characterization of a new strain of Taura syndrome virus (TSV) from Colombian shrimp farms and the implication in the selection of TSV resistant lines.

J Invertebr Pathol. 2012-9-25

[6]
New genotypes of white spot syndrome virus (WSSV) and Taura syndrome virus (TSV) from the Kingdom of Saudi Arabia.

Dis Aquat Organ. 2012-7-25

[7]
Historic emergence, impact and current status of shrimp pathogens in the Americas.

J Invertebr Pathol. 2012-3-13

[8]
Transcriptional analysis of Penaeus stylirostris densovirus genes.

Virology. 2010-4-8

[9]
Characterization of a Taura syndrome virus isolate originating from the 2004 Texas epizootic in cultured shrimp.

Arch Virol. 2010-1-5

[10]
A quick fuse and the emergence of Taura syndrome virus.

Virology. 2009-8-1

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