猪德尔塔冠状病毒实时荧光PCR检测方法的建立及初步应用

Establishment and Preliminary Application of Fluorescent PCR for Detection of Porcine Delta Coronavirus

由猪德尔塔冠状病毒(PDCoV)引起的腹泻,直接影响仔猪存活率和后期生长速度,严重的可导致死亡,是养猪企业重点监测的疫病之一。为了满足养猪企业对猪腹泻样本快速检测和排查的需要,本研究基于Taqman荧光探针技术建立了实时荧光PCR快速检测PDCoV的方法,并将其应用于猪场实际样品的检测。结果显示:本研究建立的实时荧光RT-PCR检测PDCoV的方法具有较高的特异性,可将PDCoV与同属的其他冠状病毒进行区分;其检测灵敏度比传统的PCR提高100倍;通过对大量临床样本的检测,证实该方法具有良好的稳定性,可在众多的腹泻临床样本中准确、高效地对PDCoV进行鉴定,从而减少养殖企业的损失。

Diarrhea caused by Porcine deltacoronavirus(PDCoV)directly affects the survival rate and later growth rate of piglets.In serious cases,it could lead to death.It is one of the key diseases monitored by pig breeding enterprises.In order to meet the needs of pig enterprises for rapid detection and screening of pig diarrhea samples,a real-time fluorescent RT-PCR method for rapid detection of PDCoV was established based on TaqMan probe,and applied to the detection of clinical samples in pig farms.The results showed that the real-time fluorescent PCR method established in this study had high specificity and could distinguish PDCoV from other coronaviruses of the same genus.Its detection sensitivity is 100 times higher than that of traditional PCR.For the detection of a large number of clinical samples,it is confirmed that this method has good stability,and could accurately and efficiently distinguish PDCoV in a large number of diarrhea clinical samples,so as to reduce the loss of breeding enterprises.