动物医学进展2025，Vol.46Issue(7)：18-25,8.
猪繁殖与呼吸综合征病毒三重纳米RT-PCR检测方法的建立
Development of a Nanoparticle-assisted Triplex RT-PCR Assay for Porcine Reproductive and Respiratory Syndrome Virus Detection
摘要
Abstract
Porcine reproductive and respiratory syndrome virus(PRRSV)has constantly varied and circula-ted in the pig herd causing severe economic losses worldwide.In this study,a nanoparticle-assisted RT-PCR(Nano RT-PCR)assay for PRRSV detection was established by the combination of nanoparticles with tra-ditional RT-PCR technology.PRRSV-specific general primers were designed and synthesized according to Nsp2 nucleotide sequences of PRRSV classical,highly pathogenic,and NADC30-like strains and the Nano RT-PCR method was developed after optimization of the annealing temperature,primer concentration,and the number of reaction cycles.The results of specificity test showed that no cross-reaction was observed for the Nano RT-PCR assay when detecting classical swine fever virus,porcine epidemic diarrhea virus,swine transmissible gastroenteritis virus,porcine circovirus type 2,pseudorabies virus,and porcine parvovirus.The assay was more sensitive than the conventional RT-PCR assay with detection limits of 3.32× 102,6.18× 103,and 3.26 × 103 copies/μL Nsp2 recombinant plasmid of classical,highly pathogenic,and NADC30-like strains.The detection results of 80 clinical samples showed the assay could be used for the differential detection of PRRSV classic strain,highly pathogenic strain and NADC30-like strain in clinical samples,and could determine the strain type.The assay provided technical support for the rapid detection of PRRSV.关键词
猪繁殖与呼吸综合征病毒/经典毒株/高致病性毒株/NADC30样毒株/纳米RT-PCRKey words
PRRSV/Classic strain/Highly pathogenic strain/NADC30-like strain/Nano RT-PCR分类
农业科技引用本文复制引用
茹敏,高洁,赵治钤,郑丹阳,赵谜,穆杨..猪繁殖与呼吸综合征病毒三重纳米RT-PCR检测方法的建立[J].动物医学进展,2025,46(7):18-25,8.基金项目
陕西省动物防疫专项项目(2023-33) （2023-33）
陕西省重点研发计划项目(2022NY-099) （2022NY-099）
