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安徽一株塞内卡病毒的遗传进化分析及检测方法的建立。

Genetic evolutionary analysis of a strain of Senecavirus A in Anhui and the establishment of its detection method.

机构信息

College of Animal Science and Technology, Anhui Agricultural University, Hefei, Anhui 230036, China.

Joint International Research Laboratory of Agriculture and Agri-Product Safety, The Ministry of Education of China, Yangzhou University, Yangzhou 225009, China.

出版信息

Infect Genet Evol. 2024 Oct;124:105665. doi: 10.1016/j.meegid.2024.105665. Epub 2024 Sep 2.

Abstract

BACKGROUND

Senecavirus A (SVA) is the only member of the genus Senecavirus in the family Picornaviridae, and is one of the pathogens of porcine blistering disease. SVA has been reported in the United States, Canada, China, Thailand, and Colombia.

METHODS

In this study, positive SVA infection was detected by RT-PCR in sick materials collected from pig farms of different sizes in Anhui Province.

RESULTS

In this study, a virulent strain of SVA was successfully obtained by viral isolation on BHK21 cells and named SVA-CH-AHAU-1. Meanwhile, a simple, rapid and accurate nano-PCR method for the detection of SVA infection was established in this study, using the recombinant plasmid pClone-SVA-3D as a template.

CONCLUSIONS

The complete genome of SVA-CH-AHAU-1 is 7286 bp, including a 5' non-coding region (UTR), an open reading frame (ORF) of 6546 nucleotides, encoding 2182 amino acids (aa), and a 3' UTR with Poly(A) features, and phylogenetic analysis showed that this isolate had the highest nucleotide homology (97.9 %) with the US isolate US-15-41901SD. In this study, the virulent strain SVA-CH-AHAU-1 was found to recombine in the ORF region with isolates SVA-CH-SDGT-2017 and SVA/Canada/ON/FMA-2015-0024 T2/2015. The complete genome has been submitted to GeneBank with the accession number OM654411. In addition, our results suggest that the established nano-PCR assay can be used as an economical, reliable and sensitive method for the field diagnosis of SVA method, especially in resource-limited areas.

摘要

背景

塞尼卡病毒 A(SVA)是小 RNA 病毒科鼻病毒属的唯一成员,也是猪水疱病的病原体之一。SVA 已在美国、加拿大、中国、泰国和哥伦比亚等地被报道。

方法

本研究通过 RT-PCR 检测了来自安徽省不同规模猪场的病料中的阳性 SVA 感染。

结果

本研究通过在 BHK21 细胞上进行病毒分离,成功获得了一株强毒 SVA 毒株,并将其命名为 SVA-CH-AHAU-1。同时,本研究建立了一种简单、快速、准确的纳米 PCR 检测 SVA 感染的方法,以重组质粒 pClone-SVA-3D 为模板。

结论

SVA-CH-AHAU-1 的完整基因组为 7286bp,包括 5'非编码区(UTR)、6546 个核苷酸的开放阅读框(ORF),编码 2182 个氨基酸(aa)和具有 Poly(A)特征的 3'UTR。系统进化分析显示,该分离株与美国分离株 US-15-41901SD 的核苷酸同源性最高(97.9%)。本研究发现,强毒 SVA-CH-AHAU-1 在 ORF 区与 SVA-CH-SDGT-2017 株和 SVA/Canada/ON/FMA-2015-0024T2/2015 株发生重组。该完整基因组已提交至基因库,登录号为 OM654411。此外,我们的研究结果表明,建立的纳米 PCR 检测方法可作为一种经济、可靠、敏感的 SVA 现场诊断方法,尤其适用于资源有限的地区。

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