CAJ | 학술논문

采用Vero-76细胞系从临床样品小肠内容物中分离到1株猪流行性腹泻病毒（PEDV ），通过RT-PCR进行了鉴定。针对PEDV ，参照保守的M 基因序列设计了特异性引物和探针，建立了针对PEDV特异性的实时荧光定量RT-PCR检测方法（PEDV-rRT-PCR）。所建立的方法特异性强，与传染性胃肠炎病毒、轮状病毒、猪细小病毒等猪主要病原体无交叉反应；方法灵敏度高，检测灵敏度达到重组质粒10-8稀释度（约20 cop）；重复性好，组内变异系数≤0．26％。20份田间样品检测结果表明，所建立的PEDV-rRT-PCR方法快速、灵敏，检测结果准确可靠，适合用于PEDV 的快速检测和流行病学调查。
채용Vero-76세포계종림상양품소장내용물중분리도1주저류행성복사병독（PEDV ），통과RT-PCR진행료감정。침대PEDV ，삼조보수적M 기인서렬설계료특이성인물화탐침，건립료침대PEDV특이성적실시형광정량RT-PCR검측방법（PEDV-rRT-PCR）。소건립적방법특이성강，여전염성위장염병독、륜상병독、저세소병독등저주요병원체무교차반응；방법령민도고，검측령민도체도중조질립10-8희석도（약20 cop）；중복성호，조내변이계수≤0．26％。20빈전간양품검측결과표명，소건립적PEDV-rRT-PCR방법쾌속、령민，검측결과준학가고，괄합용우PEDV 적쾌속검측화류행병학조사。
A porcine epidemic diarrhea virus (PEDV)strain was isolated with Vero-76 cell line from small intestine contents of swine with clinical signs and identified by RT-PCR assay .Primers and probe were de-signed according to the conserved M gene of PEDV ,and a Taqman real-time RT-PCR (rRT-PCR) assay was developed .The sensitivity and specificity of PEDV-rRT-PCR assay were evaluated .The specificity of rRT-PCR assays developed had no cross reaction with transmissible gastroenteritis virus ,rotavirus group A ,porcine parvovirus and others .The sensitivity of PEDV-rRT-PCR was 10-8 dilution of recombinant plasmid with about 20 copies .The assay has a good repeatability with coefficient of variation within group less than or equal to 0.26% .Twenty field samples tested with the assay indicated that PEDV-rRT-PCR was specific ,sensitive and rapid . The developed assay is an excellent method for routine detection of PEDV and could be used to monitor and survey PED .