病毒性出血性败血症病毒(VHSV)可视化核酸试纸条检测方法的建立
Establishment of a Visual Nucleic Acid Strip Test for Viral Haemorrhagic Septicaemia Virus
殷培军 1金莹 1唐静 1岳志芹 1王宫璞 1房保海1
作者信息
摘要
本研究通过在病毒性出血性败血症病毒(Viral Haemorrhagic Septicaemia Virus,VHSV)的保守区5'端设计一对保守引物,在上游和下游引物的5'端分别修饰生物素(biotin)和异硫氰酸荧光素(FITC),成功建立了VHSV快速、可视化核酸试纸条检测方法.该方法具有高灵敏度,能够检测到33拷贝/μL的VHSV,比凝胶成像检测方法提高了10倍的灵敏度.实验结果证实该方法操作程序简便、反应迅速且灵敏,具备良好的特异性,可满足现场一线对于VHSV快速检测的需求,为流行病学调查及预防控制提供了新的方法学基础.
Abstract
A rapid and visual nucleic acid test strip assay for Viral Haemorrhagic Septicaemia Virus(VHSV)was successfully established in this study.This was achieved by designing a pair of conserved primers at the 5'end of the conserved region of VHSV and modifying biotin and fluorescein isothiocyanate(FITC)at the 5'end of the upstream and downstream primers,respectively.The method has high sensitivity and is able to detect VHSV at 33 copies/μL,which is a 10-fold increase in sensitivity over the gel imaging detection method.The experimental results confirmed that the method had simple operating procedures,rapid response,sensitivity,and good specificity,which could meet the needs of the field for rapid detection of VHSV,and provided a new methodological basis for epidemiological investigation and prevention and control.
关键词
病毒性出血性败血症病毒(VHSV)/核酸试纸条/可视化检测Key words
Viral Haemorrhagic Septicaemia Virus(VHSV)/nucleic acid test strips/visual detection引用本文复制引用
基金项目
国家重点研发计划(2021YFF0602803)
出版年
2024
NETL
NSTL
万方数据