中国兽医杂志2026，Vol.62Issue(1)：45-55,11.DOI:10.20157/j.cnki.zgsyzz.2026.01.006
猪流行性腹泻病毒(GHN/HB01株)灭活病毒标准物质的研制
Development of Inactivated Virus Reference Material for Procine Epidemic Diarrhea Virus(GHN/HB01 Strain)
房琳琳 1张锋 1刘爽 1魏荣 1董雅琴 1包静月1
作者信息
- 1. 中国动物卫生与流行病学中心,山东 青岛 266200
- 折叠
摘要
Abstract
To improve the accuracy of nucleic acid detection for porcine epidemic diarrhea virus(PEDV),this study established a droplet digital PCR(ddPCR)assay for PEDV and prepared an inactivated PEDV(GHN/HB01 strain)reference material.The purity of the prepared material was evaluated by real-time fluorescent quantitative PCR(qPCR)and conventional PCR to confirm the absence of exogenous pathogens.The absolute quantification of the PEDV N gene was determined using ddPCR to assess the homogeneity and stability of the reference material,and value assignment was conducted collaboratively with nine laboratories.Furthermore,qPCR was employed to evaluate the applicability of the reference material.Results showed that the optimized PEDV ddPCR method was successfully established,and the prepared inactivated PEDV(GHN/HB01 strain)reference material was free of exogenous pathogen contamination.There were no significant differences within or between groups(P>0.05),indicating good homogeneity.The material remained stable for 9 months at-20℃,7 days at 4℃,and 24 hours at 25℃,and tolerated up to three freeze-thaw cycles without degradation.The assigned quantitative value was(1.64±0.262 4)×103 copies/µL.Application evaluation demonstrated that this reference material performed reliably in qPCR assays with excellent compatibility.In conclusion,the inactivated PEDV(GHN/HB01 strain)reference material developed in this study possesses accurate quantification and strong stability,making it suitable for use in the development of diagnostic kits,laboratory method validation,and quality control.It serves as a critical material foundation for the standardization of PEDV nucleic acid detection.关键词
猪流行性腹泻病毒(PEDV)/N基因/微滴式数字PCR(ddPCR)/标准物质/定值Key words
porcine epidemic diarrhea virus(PEDV)/N gene/droplet digital PCR(ddPCR)/reference material/value assignment分类
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房琳琳,张锋,刘爽,魏荣,董雅琴,包静月..猪流行性腹泻病毒(GHN/HB01株)灭活病毒标准物质的研制[J].中国兽医杂志,2026,62(1):45-55,11.
