鲤鱼水肿病毒实时荧光定量PCR检测方法的建立。

IF 2.2 3区 农林科学 Q2 FISHERIES
Huilin He, Ruixue Feng, Siyi Guo, Bing Han, Jinshan Huang, Xu Lin, Jiyuan Yin, Defeng Zhang, Min Liu, Wen Shi
{"title":"鲤鱼水肿病毒实时荧光定量PCR检测方法的建立。","authors":"Huilin He, Ruixue Feng, Siyi Guo, Bing Han, Jinshan Huang, Xu Lin, Jiyuan Yin, Defeng Zhang, Min Liu, Wen Shi","doi":"10.1111/jfd.70027","DOIUrl":null,"url":null,"abstract":"<p><p>Carp oedema virus disease (CEVD), caused by the carp oedema virus (CEV), results in significant economic losses to the common carp and koi aquaculture industry. The CEVD symptoms resemble those of koi herpesvirus (KHV) and include lethargy, swollen gills, sunken eyes, and skin haemorrhages. Consequently, the development of efficient detection methods is imperative for the prevention and diagnosis of CEVD. In this study, we developed a quantitative real-time PCR assay for the sensitive detection of CEV infection. Specific primers were designed based on a highly conserved region within the CEV p4a gene. The assay exhibited a sensitivity that was 100-fold greater than that of the conventional PCR technique, with high specificity, and no observed cross-reactivity with infectious haematopoietic necrosis virus (IHNV), infectious pancreatic necrosis virus (IPNV), spring viremia of carp virus (SVCV), salmonid alphavirus (SAV), or KHV. Inter- and intra-assay experiments also confirmed the method's high repeatability. In a clinical setting, it showed higher sensitivity compared to the diagnostic methods for CEVD developed by the Center for Environment, Fisheries and Aquaculture Science (CEFAS), while also significantly reducing detection costs. Consequently, the established assay could provide robust technical support for the clinical diagnosis and epidemiological investigation of CEVD.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":" ","pages":"e70027"},"PeriodicalIF":2.2000,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Development of a Sensitive Real-Time PCR Method for Detection of Carp Oedema Virus.\",\"authors\":\"Huilin He, Ruixue Feng, Siyi Guo, Bing Han, Jinshan Huang, Xu Lin, Jiyuan Yin, Defeng Zhang, Min Liu, Wen Shi\",\"doi\":\"10.1111/jfd.70027\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Carp oedema virus disease (CEVD), caused by the carp oedema virus (CEV), results in significant economic losses to the common carp and koi aquaculture industry. The CEVD symptoms resemble those of koi herpesvirus (KHV) and include lethargy, swollen gills, sunken eyes, and skin haemorrhages. Consequently, the development of efficient detection methods is imperative for the prevention and diagnosis of CEVD. In this study, we developed a quantitative real-time PCR assay for the sensitive detection of CEV infection. Specific primers were designed based on a highly conserved region within the CEV p4a gene. The assay exhibited a sensitivity that was 100-fold greater than that of the conventional PCR technique, with high specificity, and no observed cross-reactivity with infectious haematopoietic necrosis virus (IHNV), infectious pancreatic necrosis virus (IPNV), spring viremia of carp virus (SVCV), salmonid alphavirus (SAV), or KHV. Inter- and intra-assay experiments also confirmed the method's high repeatability. In a clinical setting, it showed higher sensitivity compared to the diagnostic methods for CEVD developed by the Center for Environment, Fisheries and Aquaculture Science (CEFAS), while also significantly reducing detection costs. Consequently, the established assay could provide robust technical support for the clinical diagnosis and epidemiological investigation of CEVD.</p>\",\"PeriodicalId\":15849,\"journal\":{\"name\":\"Journal of fish diseases\",\"volume\":\" \",\"pages\":\"e70027\"},\"PeriodicalIF\":2.2000,\"publicationDate\":\"2025-07-16\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of fish diseases\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://doi.org/10.1111/jfd.70027\",\"RegionNum\":3,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"FISHERIES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of fish diseases","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1111/jfd.70027","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"FISHERIES","Score":null,"Total":0}
引用次数: 0

摘要

鲤鱼水肿病毒病(Carp oedema virus disease, CEVD)是由鲤鱼水肿病毒(Carp oedema virus, CEV)引起的一种疾病,给鲤鱼和锦鲤养殖业造成了重大的经济损失。CEVD的症状与锦鲤疱疹病毒(KHV)相似,包括嗜睡、腮肿、眼睛凹陷和皮肤出血。因此,开发有效的检测方法对于预防和诊断CEVD至关重要。在这项研究中,我们开发了一种实时荧光定量PCR检测CEV感染的方法。根据CEV p4a基因的高度保守区域设计特异性引物。该检测方法的灵敏度是传统PCR技术的100倍,特异性高,且与感染性造血坏死病毒(IHNV)、感染性胰腺坏死病毒(IPNV)、鲤鱼春季病毒血症(SVCV)、鲑鱼甲病毒(SAV)或KHV无交叉反应。测定间和测定内实验也证实了该方法的高重复性。在临床环境中,与环境、渔业和水产养殖科学中心(CEFAS)开发的CEVD诊断方法相比,它显示出更高的灵敏度,同时也显著降低了检测成本。该方法可为CEVD的临床诊断和流行病学调查提供有力的技术支持。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Development of a Sensitive Real-Time PCR Method for Detection of Carp Oedema Virus.

Carp oedema virus disease (CEVD), caused by the carp oedema virus (CEV), results in significant economic losses to the common carp and koi aquaculture industry. The CEVD symptoms resemble those of koi herpesvirus (KHV) and include lethargy, swollen gills, sunken eyes, and skin haemorrhages. Consequently, the development of efficient detection methods is imperative for the prevention and diagnosis of CEVD. In this study, we developed a quantitative real-time PCR assay for the sensitive detection of CEV infection. Specific primers were designed based on a highly conserved region within the CEV p4a gene. The assay exhibited a sensitivity that was 100-fold greater than that of the conventional PCR technique, with high specificity, and no observed cross-reactivity with infectious haematopoietic necrosis virus (IHNV), infectious pancreatic necrosis virus (IPNV), spring viremia of carp virus (SVCV), salmonid alphavirus (SAV), or KHV. Inter- and intra-assay experiments also confirmed the method's high repeatability. In a clinical setting, it showed higher sensitivity compared to the diagnostic methods for CEVD developed by the Center for Environment, Fisheries and Aquaculture Science (CEFAS), while also significantly reducing detection costs. Consequently, the established assay could provide robust technical support for the clinical diagnosis and epidemiological investigation of CEVD.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Journal of fish diseases
Journal of fish diseases 农林科学-海洋与淡水生物学
CiteScore
4.60
自引率
12.00%
发文量
170
审稿时长
6 months
期刊介绍: Journal of Fish Diseases enjoys an international reputation as the medium for the exchange of information on original research into all aspects of disease in both wild and cultured fish and shellfish. Areas of interest regularly covered by the journal include: -host-pathogen relationships- studies of fish pathogens- pathophysiology- diagnostic methods- therapy- epidemiology- descriptions of new diseases
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信