基于油包水微滴技术的多重非洲猪瘟病毒核酸检测的敏感性实验OA
Sensitivity Experiment of Multiplex Nucleic Acid Detection for African Swine Fever Virus Based on Oil-in-Water Microdroplet Technology
非洲猪瘟病毒(ASFV)对全球养猪业及食品安全威胁重大.该研究利用油包水数字PCR(ddPCR)技术,开发高敏特异的多重检测方法,同步检测MGF、VP72基因及猪β-肌动蛋白内参基因.通过设计引物探针,构建阳性对照质粒并验证,优化ddPCR体系后,实现阳性微滴清晰分离.该方法敏感性高,MGF和VP72的空白值限分别为3.115拷贝/反应和1拷贝/反应,可有效区分单双阳性微滴,为ASFV检测提供可靠工具,展现了ddPCR在病毒监测及疾病防控中的应用潜力.
African Swine Fever Virus(ASFV)poses a significant threat to global pig farming and food safety.In this study,an oil-in-water digital PCR(ddPCR)technology was utilized to develop a highly sensitive and specific multiplex detection method for simultaneous detection of MGF,VP72 genes,and porcine β-actin internal reference gene.By designing primers and probes,constructing and verifying positive control plasmids,and optimizing the ddPCR system that display clear separation of positive microdroplets.The method exhibited high sensitivity,with the limit of blank values for MGF and VP72 being 3.115 copies/reaction and 1 copy/reaction,respectively.It can effectively distinguish single and double positive microdroplets,providing a reliable tool for ASFV detection and demonstrating the application potential of ddPCR in viral monitoring and disease prevention.
王海烽
黄河水利职业技术大学环境工程学院,河南 开封 475004
农业科技
非洲猪瘟病毒核酸扩增数字PCR敏感性实验
African swine fever virusnucleic acid amplificationdigital PCRsensitivity experiment
《现代畜牧科技》 2026 (5)
5-8,4
河南省科技攻关计划项目:多重数字PCR技术对非洲猪瘟痕量核酸检测体系的研发(242102310336)
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