中国猪业2025，Vol.20Issue(5)：65-72,8.DOI:10.16174/j.issn.1673-4645.2025.05.008
PDCoV N基因TaqMan荧光定量PCR检测方法的建立
Establishment of TaqMan Fluorescence Quantitative PCR Detection Method for PDCoV N Gene
摘要
Abstract
Porcine deltacoronavirus(PDCoV)is a highly pathogenic virus that causes acute watery diarrhea,vomiting and high mortality(30％～50％)in suckling piglets,posing a significant threat to the global swine industry.To develop a sensitive and accurate detection method for PDCoV,this study designed specific primers and probes targeting the conserved region of the PDCoV nucleocapsid(N)gene,optimized the reaction system,and successfully established a TaqMan-based fluorescence quantitative PCR(qPCR)assay.Us-ing a recombinant plasmid containing the PDCoV N gene as a standard,the method exhibited a strong linear correlation within the range of 8.79 × 101 to 8.79 × 108 copies/μL(R2=0.997,slope=-3.236),with optimal primer and probe concentrations of 0.4 μmol/L and 0.2 pmol/L,respectively.The assay demonstrated high sensitivity,with a detection limit of 8.79 copies/μL;excellent specificity,with no cross-reactivity observed with Porcine Epidemic Diarrhea Virus(PEDV),Pseudorabies Virus(PRV),or Porcine Reproductive and Respira-tory Syndrome Virus(PRRSV);and good reproducibility,with inter-and intra-assay coefficients of variation below 1.5％.The qPCR method developed in this study provides a reliable technical platform for the clinical diagnosis and epidemiological surveillance of PDCoV.关键词
猪/猪德尔塔冠状病毒/核衣壳基因/TaqMan qPCR/仔猪腹泻Key words
pig/PDCoV/N gene/TaqMan qPCR/piglet diarrhea分类
农业科技引用本文复制引用
马翮斯,罗灵芝,王元红,虞凌雪,李国新..PDCoV N基因TaqMan荧光定量PCR检测方法的建立[J].中国猪业,2025,20(5):65-72,8.基金项目
上海科技兴农项目腹泻二联苗研制-李国新 （）
上海市科技兴农技术创新项目(X2023-02-08-00-12-F04612) （X2023-02-08-00-12-F04612）
