The Nerve Conduction Study Of Tracing Pseudorabies Virus Infection The Anal Sphincter By Immunohistochemistry

As an important virus, Pseudorabies Virus does harm to the development of the pig industry seriously. At the same time as a new model of neural tracer study viruses, it has also been widespread concerned and researched by scientists.The study measured the virus titer of Pseudorabies SA215-T strain which constructed by our laboratory, and used the SA215-T strain infected SD rats, and established an experimentally infected rats pathological model.56 hours after infection, the rats appeared symptoms like fever, itching and ataxia and so on, about 115 hours begin to die, anatomical lesions were pulmonary edema, diffuse brain hemorrhage. The rats’mean survival time after infection PRV SA215-T were 120 hours, compared with the mean survival time of infected PRV Bartha-K61 strain’s average survival time 114 hours, a litter longer, so PRV SA215-T is suitable for the establishment of PRV infected rats pathological model.In this study were used the PRV SA215-T and Bartha-K61 artificially infected rats, observed and recorded the clinical symptoms and survival situation after rats infected with virus. Dissected the rats at different times and collected liver, spleen, lung, and kidney and brain tissues and organs. Then detected the PRV antigen’s distribution organizations of the organ in rats by PCR method. The results showed that rats infected with PRV within 48 hours after the tissues and organs within the PRV antigen was not detected; About 56 hours after infection, PRV antigen was positive distribution in lung; Over time, the PRV antigen distributed in tissues and organs was more widely, ultimately almost throughout all tissues and organs. This study established an indirect immunohistochemistry method for detected PRV antigen and optimized the immunohistochemical criteria. By this immunohistochemical method can detected the PRV antigen in the tissues and organs of rats. The method for clinical testing was simple, sensitive, specific, staining easy to observe.In this study, detected and located the PRV SA215-T’s position in the spinal cord after infection by the indirect immunohistochemical method which established before, tracing the nerve pathways of PRV infected in anal sphincter of rats:the first time detected PRV antigen in spinal cord lumbar segments 4-5 after infection was 48 hours; about 56 hours after infection, thoracic 11-12 segments detected PRV antigen; about 66 hours after infection in spinal cord cervical vertebra segments 6-7 detected PRV antigen; about 72 hours after infection, in the brain detected PRV antigen. After gene deletion, pseudorabies virus has reverse nerve conduction characteristics, using immunohistochemical methods can specifically track its pathway in the spinal nerve. In this study, used the pathological model which PRV infected in anal sphincter of rats, proved the anal sphincter nerve pathways, laid the foundation for further study the pathogenesis and developing laws of congenital anal atresia disease.