| This study concentrates on①developing the Real-time PCR method for GPV-VP3 detection,②studing the dynamic distribution of GPV-VP3 gene vaccine (pcDNA-GPV-VP3)in geese (duodenum, liver, spleen, kidney, fabricius, thymus, ileum, gland of Harder, position of injected spot, rectum, cecum, pancreas, heart, blood, jejunum, lung, and brain), through 30-day-old geese were injected pcDNA-GPV-VP3 with different doses via gene-gun (6, 3, 1μg), intramuscular injection (200μg, 100μg, 50μg) respectively and live attenuated vaccine of GPV(200μL),③studing the dynamic distribution of live attenuated vaccine of Sichuan isolate of GPV.Results showed that:1. This assay was specific, highly sensitive, and rapid for detecting pcDNA-GPV-VP3. The standard curve showed a fine linear relationship between Ct and template concentration, and the correlation coefficient was 0.999.2. Dynamic distribution of GPV-VP3 gene vaccine in gosling: The pcDNA-GPV-VP3 could be detected in all tissues and peripheral blood at 1h post infection, the copy number was most at the position of injected spot, then liver, kidney, lymphatic organ (spleen, fabricius, thymus, gland of Harder); At 217d post administration, pcDNA-GPV-VP3 could still be detected in tissues of different doses group. But the copy number decreased about 103-104 copies in the most tissues than it at 1h post administration, the obvious decrease was found at the injection sites (it dropped about 104-105 copies); The changement of the copy numbers in the serum was always little and at 1h post administration was less about 104-105 copies than other tissues; then the copy numbers decreased, with the time passed slowly, decreased 100.1-100.5 in 3d-21d, only 100.5-101 in 63d-217d; in brain the copy number increased in 1h-12h, reach the peak at 12h, then deceased.3. Dynamic expression of GPV-VP3 gene vaccine in gosling: The expression of pcDNA-GPV-VP3 could be detected at 1d post infection from the cytoplasm of injected spot, the positive numbe reached peak at 1d via vaccined with gene gun bombardment, and at 7d with intramuscular injection. At 63d post administration, expression could still be detected in the sarcoplasmand of injected spot, it also could be detected easier in cellular of cardiac muscle, caliciform cell in villi intertinales and gl. Intersinales, intercellular substance of liver; other tissues had no expression.4. Different dynamic distribution and expression of GPV-VP3 gene vaccine in gosling via the different Immunogenicity: pcDNA-GPV-VP3 DNA vaccine immunized geese respectively by the different routes of gene gun bombardment and intramuscular injection, which all could be detected in all tissues, and expressed VP3 in liver, ileum, cecum, heart, jejunum, duodenum, rectum, position of injected spot; pcDNA-GPV-VP3 DNA immunized by gene gun bombard- ment demonstrated higher copy numbers in all tissues and earlier expressed in liver, cecum than intramuscular injection at nonage; but at 217d post administration were no different.5. Different dynamic distribution and expression of GPV-VP3 gene vaccine in gosling with the different doses: The quantity of pcDNA-GPV-VP3 and expression detected in the experimental groups showed the relations with the quantity of the inoculation doses, but not proportion (P≥0.05).6. Infection and excretion of attenuated vaccine of GPV in gosling: Geese were immunized with live attenuated vaccineof GPV, the copy number decreased at the beginning then increased in 1h-3d, the maximum amount at 7d post administration, after 35d the copy number decreased gradually; At the position of injected spot, heart, liver and intestinal, viral antigens were detected in 1d-3d, the maximum amount at 7d; compared with the DNA vaccine, attenuated GPV vaccine was detected more copy number and expression in tissues, the difference was significant (P≤0.01≤0.05); but the number dropped quickly, all tissues failed to be detected antigen at 63d.
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