福建农业学报2024，Vol.39Issue(10)：1146-1151,6.DOI:10.19303/j.issn.1008-0384.2024.10.005
猪流行性腹泻病毒荧光RT-RAA检测方法的建立及初步应用
Fluorescent RT-RAA for Diagnosing Epidemic Diarrhea in Pigs
摘要
Abstract
[Objective]A rapid fluorescence RT-RAA to detect porcine epidemic diarrhea virus(PEDV)was developed and tested for field application.[Methods]Primers and probes were designed for the conserved region of PEDV-S gene fragment,and a standard plasmid was constructed.Through condition optimization followed by tests for assay specificity,sensitivity,and repeatability,a recombinant enzyme-mediated chain replacement nucleic acid amplification fluorescence RT-RAA for detecting PEDV was established.[Results]Under a constant temperature of 42℃for 20 min,the assay detected PEDV as positive,while negative on the porcine transmissible gastroenteritis virus(TGEV),classical swine fever virus(CSFV),porcine pseudorabies virus(PRV),porcine reproductive and respiratory syndrome virus(PRRSV),porcine rotavirus(PoRV)and other porcine viruses.It had a minimum detection limit of 4.43×102 standard plasmid copies·μL-1,a reproducibility showing little difference between the standard plasmids of the same concentration,and a positive rate of 7.5%(3 out of 40)on PEDV specimens comparable to that obtained by RT-qPCR.[Conclusion]The newly developed fluorescence RT-RAA for PEDV detection was considered appropriate for rapid diagnosis of epidemic diarrhea in pigs.关键词
猪流行性腹泻病毒/S蛋白/RT-RAAKey words
porcine epidemic diarrhea virus/S protein/fluorescence RT-RAA分类
农业科技引用本文复制引用
侯恩慧,陈秋勇,吴学敏,丘镜莉,吴仁杰,周伦江,刘玉涛,马玉芳,王隆柏..猪流行性腹泻病毒荧光RT-RAA检测方法的建立及初步应用[J].福建农业学报,2024,39(10):1146-1151,6.基金项目
福建省科技计划公益类专项(2021R10260016) （2021R10260016）
福建省科技重大专项(2021NZ029023) （2021NZ029023）
福建省农业高质量发展超越"5511"协同创新工程项目(XTCXGC2021008) （XTCXGC2021008）
