摘要
猪德尔塔冠状病毒(porcine delta coronavirus,PDCoV)是2012年中国香港首次发现的冠状病毒,能引起感染仔猪水样腹泻,最终脱水死亡。研究根据PDCoV保守的衣壳蛋白基因(N)序列设计特异性引物,建立荧光定量PCR检测方法。结果表明,该方法在6.31×10^6~6.31×10^1copies/μL,Ct与模板浓度之间具有良好的线性关系,斜率为-4.90,相关系数为0.999。所有标准品模板出现唯一熔解峰,敏感性下限为6.31×10^1copies/μL,PEDV和TGEV等均没有检测信号。对60份临床样品检测,PDCoV阳性率为5.0%,表明本研究建立的方法可以快速灵敏地检测PDCoV,从而为PDCoV的预防和疫苗研制奠定基础。
Porcine delta coronavirus(PDCoV)is a newly emerging enterotropic swine coronavirus that causes acute enteritis in nursing piglets.A sensitive real-time PCR method is urgently required to detect PDCoV infection.In this study,we designed and synthesized specific primers based on the conserved sequence of PDCoV N gene to construct a standard plasmid.The results showed that the slope of the standard curve of PDCoV real-time fluorescence quantitative detection method was-4.90,and the correlation coefficient was 0.999.Specific melting peaks appeared in all dilution standard templates;using 10-fold gradient dilution of plasmid standards,a minimum detection was 6.31×10^1 copies/μL of plasmid DNA,demonstrating that the method has good sensitivity;No signal was detected when PEDV and TGEV were used as templates.60clinical pig feces samples were tested and the results showed that the PDCoV positive rate was 5.0%by this method.The method established in this study is suitable for the rapid detection of PDCoV in clinical samples,which could differentiate PDCoV from PEDV and TGEV.
作者
张世亨
黄海鑫
尹彦文
施开创
郑敏
汪伟
曹亮
赵翠青
刘立明
鲁会军
孙文超
金宁一
ZHANG Shi-heng;HUANG Hai-xin;YIN Yan-wen;SHI Kai-chuang;ZHNEG Min;WANG Wei;CAO Liang;ZHAO Cui-qing;LIU Li-ming;LU Hui-jun;SUN Wen-chao;JIN Ningyi(College of Animal Science and Technology,Jinlin Agricultural University,Changchun 130118,China;Institute of Virology,Wenzhou University,Wenzhou,Zhejiang325035,China;Guangxi Center for Animal Disease Control and Prevention,Nanning 530001,China;Institute of Military Veterinary,Academy of Military Medical Sciences,Changchun 130122,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2019年第12期2288-2292,共5页
Chinese Journal of Veterinary Science
基金
国家重点研发计划资助项目(2018YFD0500104)
浙江省青年基金资助项目(LQ19C180001)
温州市基础性科研基金资助项目(N20180010)
广西科技重大专项资助项目(桂科AA17204057)
广西水产畜牧科技资助项目(桂渔牧科201528017)
