Studies On The Distribution Of Gosling Parvovirus In Infected Gosling By Immunoperoxidase Staining And PCR Method

This study established successfully indirect immunoperoxidase staining method to examine goose parvovirus (GPV) distribution in the infected goslings by repeated optimization of reaction condition, including incubation time and concentrations of 3% H2O2, antigen retrieval method, antibody dilution and incubation time. After that, clinical trial, absorption experiment, substitution experiment and positive, negative control experiment indicated that it had the merits of high sensitivity and specificity. For the PCR method, primers were designed to amplify GPV NS2 gene specifically, which based on the complete nucleotide sequence of goose parvovirus submitted to the Genebank with the software of Oligo 6.0. The merits of indicating the definite position of viral antigen with immunoperoxidase staining method and high sensitivity and specificity of PCR method were combined. Furthermore, the viral antigen intensity of immunoperoxidase staining specimen was analyzed with image analysis software. In this article, the dynamic distribution of parvovirus antigen in organs and tissues of infected goslings at different time were studied.The examination result indicated that GPV was found in heart, lungs, spleen, bursa of Fabricius, thymus, proventriculus, duodenum, jejunum, ileum and blood 1d postinfection (PI). Then, liver and cecum showed GPV existence 3d PI besides the tissues montioned above. Brain, excrement and cloacal swab started to be able to examine GPV 4d PI. At 5d PI, GPV was found in esophagus. At this time, GPV was most widespread; since 6d PI, positive tissures were less, meanwhile GPV could not be found in the liver and cloacal swab. At 7d PI, GPV could not be found in the liver, caecum and the blood. Moreover, positive tissues reduced obviously 14d PI, GPV was only found in jejunum, thymus and esophagus. At 21d PI, GPV was only found in jejunum. However, GPV was not found in pancreatic gland and the skeletal muscle throughout experimental time.In generaly, GPV distributed widely in hepatocytes, the intestinal tract epithelial cells, the intestinal glands cells, the pulmonary alveolus wall cells and renal tubular epithelial cells. Obviously the epithelial cells were the main target cells of GPV. In the immunity organ, GPV could be demonstrated in the spleen, bursa of Fabricius and thymus, and thymus was more obvious. It was found that antigen reactivity was the most significantly during 4~6d PI through quantitatively analysis of the antigen intensity of immunoperoxidase staining positive tissures. Furthermore, the intensity of GPV antigen in jejunum and thymus was observed was higher compared with those of other tissues. These further proven that jejunum might be possibly GPV's target organ and the immune system (especially thymus) was markedly damaged.GPV distribution regularity in the tissues and organs of GPV-infected goslings were studied qualitatively and quantitatively in this experiment, and it certainly provide scientific basis to further study the pathogenesis mechanism and prepare the prophylactic measures of GPV infection.