文献类型：期刊文献

中文题名：猪瘟病毒在猪体组织中的分布

英文题名：Tissue Tropism of Classical Swine Fever Virus Using TaqMan qPCR Method

作者：张秋雨[1];李鹏[2];陈晴[1];任鹏举[3];银梅[1];王选年[2]

第一作者：张秋雨

机构：[1]河南科技学院动物科技学院;[2]新乡学院生物技术研究中心/生命科学与技术学院;[3]郑州大学生命科学学院

第一机构：河南科技学院动物科技学院,河南新乡453003

年份：2018

卷号：39

期号：7

起止页码：6-12

中文期刊名：动物医学进展

外文期刊名：Progress in Veterinary Medicine

收录：CSTPCD;;北大核心:【北大核心2017】；

基金：国家自然科学基金项目(31201877);国家重点研发计划项目(2016YFD0500702)

语种：中文

中文关键词：猪瘟病毒;E0基因;Taq;Man实时荧光定量PCR;组织分布

外文关键词：Classical swine fever virus;E0 gene;Taq Man qPCR;tissue distribution

摘要：建立一种猪瘟病毒TaqMan qPCR检测方法,为研究急性死亡期猪瘟病毒在组织中的分布提供一种可靠的方法。采用病理变化和实验室诊断方法对河南新乡某种猪场送检的病死猪进行诊断。根据猪瘟病毒特异性靶基因E0,设计并合成1对引物和水解探针,建立了TaqMan qPCR方法。以不同浓度的重组质粒pMD19-T-E0为模板,生成标准曲线,并对其重复性和特异性进行了验证,并用建立的方法对该病死猪的内脏组织进行检测,分析猪瘟病毒在体内组织中的差异性。利用建立的猪瘟病毒TaqMan qPCR检测方法,从PPV、PCV和PEDV等基因组的cDNA/DNA中不能扩增出目的条带;对3个不同的浓度梯度的重组质粒进行检测,其批间、批内的变异系数均小于2%,表明该方法的特异性和重复性较好。对猪瘟内脏组织病料进行检测发现,其猪瘟病毒主要集中在淋巴结、脾脏、肠道组织、血液及心脏等组织中,不同组织器官中的病毒载量也存在着一定的差异。该研究对疑似感染猪瘟病毒急性死亡的病猪进行了鉴别诊断,同时用建立的TaqMan qPCR方法检测了猪瘟病毒在组织中分布的差异性,为猪瘟的科学防控及发病机制的研究奠定了基础。
This study established a method for the detection of classical swine fever virus(CSFV)by Taq Man qPCR to provide a reliable method for the study of the distribution of CSFV in tissues of acute stage of death.The pathological lesions and laboratory diagnosis methods were used for diagnosis of a dead pig from a farm in Henan Xinxiang.A pair of primers and hydrolysis probes were designed and synthesized according to the specific target gene E0 of CSFV,and the Taq Man qPCR method was established.The standard curve of Taq Man qPCR method was established by using different concentrations of recombinant plasmid pMD19-T-E0 as template,and its reproducibility and specificity were verified.The CSFV in internal organs were detected.Taq Man qPCR method for detection of CSFV was established,which cannot amplifiy the specific bands from PPV,PCV and PEDV;3 different concentration gradient detections showed that the coefficients of variation of the inter or intra batches were all less than 2%,indicating that the method had good specificity and repeatability.The CSFV mainly concentrated in lymph node,spleen,intestine,blood and heart tissues,the viral loads in different organs were different.The study made the differential diagnosis of suspected swine fever virus infection of acute dead pigs,and used the Taq Man qPCR method to detect the CSFV distribution in tissues,which provided theoretical foundation and technical support for scientific research,prevention and control of CSFV outbreak and pathogenesis.