中国兽医学报2024,Vol.44Issue(8) :1585-1592.DOI:10.16303/j.cnki.1005-4545.2024.08.02

非洲猪瘟病毒核酸可视化检测方法的建立与评价

Establishment and assessment of a visual detection method for nucleic acid of Afri-can swine fever virus

刘星岐 白玉洁 张梦瑶 黄静波 刘光亮 李媛媛 谭树义 张海丽 张艳 曹宗喜 王化磊 黄培
中国兽医学报2024,Vol.44Issue(8) :1585-1592.DOI:10.16303/j.cnki.1005-4545.2024.08.02

非洲猪瘟病毒核酸可视化检测方法的建立与评价

Establishment and assessment of a visual detection method for nucleic acid of Afri-can swine fever virus

刘星岐 1白玉洁 1张梦瑶 1黄静波 1刘光亮 2李媛媛 1谭树义 2张海丽 1张艳 2曹宗喜 2王化磊 1黄培1
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作者信息

  • 1. 吉林大学动物医学学院人畜共患传染病重症诊治全国重点实验室,吉林长春 130062
  • 2. 海南省农业科学院畜牧兽医研究所海南省热带动物繁育与疫病研究重点实验室,海南海口 571100
  • 折叠

摘要

非洲猪瘟(African swine fever,ASF)是由非洲猪瘟病毒(African swine fever virus,ASFV)引起猪的一种高度传染性病毒性疾病.本病传染性强、致病性高,自2018年以来在我国迅速蔓延流行,对我国生猪养殖业造成了巨大的影响.为实现ASFV的快速检测,本研究将环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)与封闭式核酸可视化试纸相结合,建立了一种检测ASFV B646L基因的核酸可视化方法.该方法在65 ℃条件下,50 min内可检出1.16 copies/μL的重组质粒,且该方法与猪瘟病毒、猪繁殖和呼吸综合征病毒、猪细小病毒、猪传染性胃肠炎病毒无交叉反应.此外该方法无需特殊设备,操作简便,能有效避免传统等温扩增气溶胶污染造成假阳性的缺点.本研究为ASF疑似感染病例的筛选与早期诊断提供了一种快速、便捷、灵敏、可靠的候选方法.

Abstract

African swine fever(ASF),caused by the African swine fever virus(ASFV),is a highly contagious infectious disease of pigs.This disease has been spread rapidly in China since 2018,po-sing a huge threat to China's pig farming industry.To rapid detect the ASFV,a loop-mediated iso-thermal amplification(LAMP)combined with the disposable nucleic acid visualization test strip was established for visual detection of the nucleic acid of ASFV B646L gene.The method was easy to operate without special instruments and equipment,while it effectively avoided the disadvantage of false positives caused by aerosol contamination.The method was able to detect 1.16 copies/µL of the recombinant plasmid in 50 min at 65 ℃.In addition,the method was specific with no cross-re-action with classical swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,transmissible gastroenteritis virus.The results in this study provides a rapid,con-venient,sensitive and reliable method for early diagnosis and screening for ASFV suspected infec-tion cases.

关键词

非洲猪瘟病毒/核酸检测/环介导等温扩增/可视化检测

Key words

African swine fever virus/nucleic acid test/loop-mediated isothermal amplification/visu-al detection

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基金项目

海南省院士创新平台专项基金资助项目(YSPTZX202014)

出版年

2024
中国兽医学报
吉林大学

中国兽医学报

CSTPCDCSCD北大核心
影响因子:0.702
ISSN:1005-4545
参考文献量15
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