【摘要】 【目的】建立一种快捷、灵敏、简便检测猪流行性腹泻病毒（porcine epidemic diarrhea virus,PEDV）的RT-RAA检测方法，以提高猪流行性腹泻病毒临床检测效率。【方法】针对PEDV S基因片段保守区设计引物和探针，构建标准质粒PEDV-S，通过特异性、敏感性、重复性测定及条件优化，建立检测PEDV重组酶介导链置换核酸扩增荧光法（RT-RAA）。【结果】在42℃恒温作用20 min的条件下，建立的检测方法对检测猪传染性胃肠炎病毒（porcine transmissible gastroenteritis virus,TGEV）、猪瘟病毒（classical swine fever virus,CSFV）、伪狂犬病病毒（porcine pseudorabies virus,PRV）、猪繁殖与呼吸综合征病毒（porcine reproductive and respiratory syndrome virus,PRRSV）和猪轮状病毒（porcine rotavirus,PoRV）等猪源病毒均为阴性，猪流行性腹泻病毒（PEDV）为阳性；最低检出限为4.43×10²拷贝·μL^-1的标准质粒；重复性结果显示，相同浓度标准质粒的差异性很小；利用建立的RT-RAA方法检测40份猪病毒样本，阳性率为7.5%（3/40），与实时荧光定量PCR检测结果相同。【结论】本研究建立的PEDV RT-RAA检测方法具有快速简便、耗时短、特异性高、灵敏度强和重复性好等特点，适用于对PEDV的快速诊断。更多还原

【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.更多还原