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Study on Dynamic Distribution in Ducks and the Ablity to Anti Duck Plague Virus with Expressive Product in Vitro of DuIFN-α Gene Vaccine

Author: DuanKun
Tutor: WangMingShu;ChengAnChun
School: Sichuan Agricultural University
Course: Preventive Veterinary Medicine
Keywords: Real-time PCR The Ap alpha - interferon DNA vaccine Gene gun Dynamic Distribution Anti-virus capabilities Duck plague
CLC: S858.32
Type: Master's thesis
Year: 2007
Downloads: 69
Quote: 2
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Abstract


28-day-old duck by intramuscular injection of 50 ug, 100 ug and 200 ug and gene gun bombardment immune 1μg 3μg and 6μg duck alpha-interferon DNA vaccine (pcDNA the-SDIFN-alpha), located PBS and empty vector pcDNA3.1 () control. Duck serum and whole blood and various parts of the organization (heart, liver, spleen, lung, kidney, bursa of Fabricius, thymus, Harderian gland, pancreas, duodenum, jejunum, ileum and rectum were collected at different times after immunization , the cecum, brain and immune injection site Organization). The following studies: (1) the establishment of a detection the the pcDNA-SDIFN-alpha of real-time quantitative PCR (FQ-PCR) method and applied to the detection the the pcDNA-SDIFN-alpha in immune duck vivo dynamic distribution. Expression product (serum) anti-duck plague caused by virulent duck fibroblasts lesions the ② pcDNA-SDIFN-alpha immune duck in vivo by different doses of different ways. The results obtained are as follows: 1. Established FQ-PCR detection the pcDNA-SDIFN-alpha method has good specificity, reproducibility, wide linear range of detection, detected 7.7 × 10 to 1 to 7.7 × 10 9copies/μL, can be used to duck vivo the pcDNA-SDIFN-alpha for quantitative detection. 2. Immune duck through intramuscular injection and gene gun bombardment organizations 1h after immunization detected the distribution of the the pcDNA-SDIFN-alpha, and the highest concentrations organizations are at the immunization site, including intramuscular immune 50μg, 100μg and 200 microg group the highest copy number 9.1 × 10 7copies / g, 1.48 × 10 8copies / g, and 1.78 × 10 ~ 8; the gene gun immunization 1μg, 3μg and 6μg the highest copy number, respectively, 1.0 × 10 to 8copies / g , 2.51 × 10 ~ 8copies / g and 3.02 × 10 to 8copies / g. The intramuscular immunization bombardment immune are good immune. 3. In addition to the blood, with the immune passage of time, the pcDNA-SDIFN-alpha levels in the organization to achieve the highest after gradually decreasing trend during the immune ago 7d reduce the most obvious. 210d addition to the part of the organization is still able to detect the presence of nucleic acid vaccines, but the content is very low, only about 10 to 1 the the pcDNA-SDIFN-alpha copy number per gram of tissue. PcDNA-SDIFN-α content in whole blood has been lower in the 1H-105d, overall also showed a decreasing trend, but differences in copy number changes do not significantly significantly (P> 0.05); 210D when other organizations of the content difference is not significant with (P> 0.05). Throughout the detection period, the immunization site the pcDNA-SDIFN-alpha content reduced maximum reduction of approximately 6-7 orders of magnitude, and other tissues also reduces the content of approximately 2-5 orders of magnitude. 4. Different dose intramuscular injection of immunohistochemistry and gene gun bombardment immunohistochemistry various tissues and organs in the pcDNA-SDIFN-alpha content presented general rules are: muscle group: 200μg group> 100 ug group> 50 ug group; gene gun group: 6μg group> group 3μg > 1μg group, but the differences were not significant (P> 0.05). 5. PBS control and empty vector pcDNA3.1 () of various tissues and organs of the control group at all time points were not detected to the pcDNA-SDIFN-alpha. 6. 1D-63d after immunization, expression product acts on the duck embryo fibroblasts, alpha test group of pcDNA-SDIFN-duck plague virus levels were lower than PBS and empty vector virus content of the control group, and in the days after immunization 21d expression product role the DPV virus content than the control group, the largest reduction, indicating that the ability to inhibit the virus after immunization 3d to 21d when the expression product is the most obvious, the maximum inhibition of the cells of the virus approximately 10 to 8copies/100μL of. Intramuscular group 7d expression product of the highest inhibition of duck plague virulent gene gun immunohistochemistry virulent duck plague the strongest inhibition of the expression product 3d. 7. Different doses of intramuscular immunohistochemistry and gene gun bombardment immunohistochemical expression product inhibition of viral capacity: muscle group: 200μg group> 100 ug group> 50 ug group; gene gun group: 6μg group> 3μg> 1μg group. Indicating that the amount of expression product positively correlated with the dose of DNA vaccine immunization in the same manner. The intramuscular group of 8.200μg with 6μg gene gun immunohistochemistry organizations in the pcDNA-SDIFN-alpha content difference not significant (P> 0.05); the 6μg bombardment immunohistochemistry and 200 microg expression product inhibition of viral ability is quite 1μg bombardment immunohistochemistry and 50 ug intramuscular group expression products inhibit viral ability, indicating that the gene gun immunization is about 40 times higher than intramuscular immunization efficiency.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Livestock, poultry, wildlife diseases > Poultry > Duck
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