猪血凝性脑脊髓炎病毒在人工感染仔猪体内侵染过程和分布规律的研究  

The invading process and distribution of swine hemagglutinating encephalomyelitis virus in artificially infected piglets

作　　者：王栋[1] 臧德跃[1] 赵魁[1] 张冰冰[1] 宋德光[1] 贺文琦[1] 陈克研[1] 高丰[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《中国预防兽医学报》2011年第2期89-92,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30871849)

摘　　要：为研究血凝性脑脊髓炎病毒(HEV)在人工感染仔猪体内的侵袭过程和分布规律,本实验采用滴鼻的方式感染5头1日龄未食初乳仔猪,并在感染后每隔24 h迫杀1头仔猪,采集鼻黏膜、气管、口腔黏膜、舌、食管、胃、肠、心肌、肝、脾、肺、肾、各段脊髓和脑等组织脏器制作切片,通过间接免疫组织化学方法检测HEV在仔猪体内的动态侵袭过程。此外,采用SYBR GreenⅠ实时荧光定量PCR(Real-time PCR)方法检测病毒在各组织脏器中的分布情况。结果显示:感染病毒后2 d～3 d,仔猪出现精神沉郁、全身震颤等中枢神经系统症状,免疫组织化学检测可见病毒抗原首先出现于脑桥,并进一步蔓延至延髓和各段脊髓,最后进入小脑浦肯野氏细胞和大脑皮层锥体细胞中;Real-time PCR检测结果显示病毒广泛分布于大脑皮层、脑桥、延髓、脊髓和小脑等中枢神经系统的组织中,其中大脑皮层的检出率最高。To investigate the invading process and distribution of swine hemagglutinating encephalomyelitis virus（HEV） in artificially infected piglets,five colostrum-deprived pigs were inoculated with HEV strain 67N at the day of birth by the intranasal route and sacrificed every 24 hours postinfection;Tissue samples from mucous membrane of nose,trachea,mucous membrane of mouth,tongue,esophagus,stomach,intestine,heart,liver,spleen,ung,kidney,spinal cord and brain were collected and paraffin slices of these samples were prepared.The method of immunohistochemical staining（IHC） was applied to investigate the invading process of virus antigen;A SYBR GreenⅠreal-time PCR method was used to evaluate the distribution of viral RNA in the tissues.The results revealed that central nervous system symptoms like body trembles and spiritual depressed were observed after an incubation period of 2 to 3 days,the result of IHC showed that virus antigen were first detected in the neurons of the pons,then spreaded into medulla oblongata and spinal cord,at last the virus was found in the the purkinje cells of the cerebellum and pyramidal cells of the cerebral cortex;The result of SYBR GreenⅠreal-time PCR showed that the virus widely distributed in the tissues of cerebral cortex,pons,medulla oblongata,spinal cord and cerebellum,especially found regularly in cerebral cortex.

关 键 词：猪血凝性脑脊髓炎病毒 免疫组织化学 SYBRGreenⅠ实时荧光定量PCR 

分 类 号：S852.65[农业科学—基础兽医学]