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.

中文翻译：

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从存档的戴维森固定石蜡包埋的虾组织中检出Taura综合征病毒并对其进行系统发育分析。

Taura综合征是世界动物卫生组织（OIE）列出的海洋虾疾病，由Taura综合征病毒（TSV）（一种单链RNA病毒）引起。在这里，我们演示了使用15岁的戴维森固定石蜡包埋（DFPE）虾组织存档进行TSV检测和系统发育分析的实用程序。使用三种市售试剂盒从已知的被TSV感染的DFPE组织中分离总RNA，并比较分离出的RNA中TSV的纯度和检测能力。通过RT-qPCR在所有测试样品中成功检测到TSV。通过RT-PCR筛选的TSV特异性引物中，RNA依赖性RNA聚合酶（RdRp）的引物对TSV-20，衣壳蛋白基因VP2的引物TSV-15和TSV-16，衣壳的引物TSV-5。蛋白质基因VP1扩增的样本数量最多。为了评估不同TSV分离株之间的系统发育关系，对VP1基因进行了扩增，并在重叠片段中进行了测序。将较小片段的串联序列用于系统发育分析。结果表明，该研究的TSV分离株通常与相应地理区域的同源分离株聚集在一起，表明源自DFPE组织的RNA可用于病原体检测和回顾性分析。从存档的组织中进行基因组表征的能力将加快病原体的发现，诊断工具的开发，并防止疾病在全世界虾类和潜在的其他水产养殖物种中传播。将较小片段的串联序列用于系统发育分析。结果表明，该研究的TSV分离株通常与相应地理区域的同源分离株聚集在一起，表明源自DFPE组织的RNA可用于病原体检测和回顾性分析。从存档的组织中进行基因组表征的能力将加快病原体的发现，诊断工具的开发，并防止疾病在全世界虾类和潜在的其他水产养殖物种中传播。将较小片段的串联序列用于系统发育分析。结果表明，该研究的TSV分离株通常与相应地理区域的同源分离株聚集在一起，表明源自DFPE组织的RNA可用于病原体检测和回顾性分析。从存档的组织中进行基因组表征的能力将加快病原体的发现，诊断工具的开发，并防止疾病在全世界虾类和潜在的其他水产养殖物种中传播。