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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

Posted on:2008-03-29Degree:MasterType:Thesis
Country:ChinaCandidate:K DuanFull Text:PDF
GTID:2143360218954440Subject:Prevention of Veterinary Medicine
Abstract/Summary:PDF Full Text Request
The 28-day beijing ducks were injected intramuscularly DuIFN-αgene vaccine ( pcDNA-SDIFN-α) with 50μg, 100μg and 200μg and 1μg, 3μg and 6μg by gene gun respectively, with PBS and pcDNA3.1 (+) as controls.Gather peripheral blood and the organs (heart, liver, spleen, lung, kidney, brain, thymus gland, harderian gland, bursa of fabricius, pancreas, duodenum, jejunum, ileum, rectum, appendices and muscle at the inoculation site) at different time site post inoculation,The objective of the study is as follows:①Real-Time Fluorescent Quantitative PCR (RQ-PCR) was established and applied to detect the dynamic distribution in ducks.②Study on the ability to anti duck plague virus with expressive product in vitro of pcDNA-SDIFN-αwhich immured with different doges and different ways with TaqMan-MGB PCR. The results showed that:1 FQ-PCR method for detecting pcDNA-SDIFN-αwas established, this assay was specific, highly sensitive, and 7.7×101~7.7×109 copies/μL could be detected.This assay could be used to detect pcDNA-SDIFN-αin ducks in vitro.2 The pcDNA-SDIFN-αcould be detected in all tissues and peripheral blood at 1 hour post infections (hpi) . The most copy numbers were detected at the injection sites (P≤0.05) and the copies numbers were 9.1×107,1.48×108,1.78×108 in 50μg,100μg and 200μg immured by intramuscular injection,and 1.0×108 copies/g,2.51×108 copies/g,3.02×108 copies/g in 1μg,3μg and 6μg groups respectively immured by gene gun.It was nice way by intramuscular injection and gene gun.3. Except in peripheral blood, the copies numbers in each tissue increased firstly and then decreased with time-elapsed, and the decrement was obvious in 7dpi. The pcDNA-SDIFN-αcould still be detected in tissues and peripheral blood 210d post administration, however the numbers were a little, and The copies numbers was only about 101 copies/g.The copy numbers of the pcDNA-SDIFN-αin the serum were lower than the rest tissues, the increase and the decrease was not obvious, hower the copies numbers was declined also, but the difference between other tissues of copy numbers was not obviously at 210dpi (P>0.05) . At 1hour to 210 day, the copies numbers of the pcDNA-SDIFN-αat the injection sites decreased remarkably and it dropped about 106-7 copies/g, others also dropped about 102-5 copies/g.4. The chief rule of the pcDNA-SDIFN-αcontent in different tissues of the ducks which immunized with different doses and different way was: in 200μg groups>in100μg groups>in 50μg groups by intramuscular injection, and in 6μg groups>in 3μg groups>in 1μg groups by gene gun. However, it was not significantly differentia (P>0.05).5. The pcDNA-SDIFN-αwas not be detected in PBS group and pcDNA3.1 (+) group in all time sites.6. The DPV copies numbers in pcDNA-SDIFN-αgroups were all lower than PBS group at 1-63day post immunization (dpi) , and the ability of anti-DPV was obviously at 3-21 dpi, it was about 108copies/100μL.The ability of anti-DPV was homology between the group of 6μg and the group of 200μg.Hower the difference was that the strong ability of anti-DPV site was 7dpi in 200μg groups but in 3dpi in 6μg groups7. The ability of anti-DPV of the espression products was that 200μg>100μg>50μg in the group by intramuscular injection and 6μg>3μg>1μg by gene gun, it showed that there was some correlation between the quantity of expression product and the dosage8. The difference of cDNA-SDIFN-αcopies numbers was not obvious between in 200μg and 6μg, and the ability of anti-DPV by gene gun was equal to intramuscular injection, So that the matter by gene gun was better than intramuscular injection, and the efficiency was 40 times lager than the manner by intramuscular injection.
Keywords/Search Tags:FQ-PCR, SDIFN-α, gene vaccine, Gene gun, bio-distribution, anti-viral ability, DPV
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