5 results about "Duck circovirus" patented technology

The invention discloses a duck circovirus ELISA antibody detection kit and application thereof, and relates to the technical field of biological detection. The duck circovirus ELISA antibody detection kit comprises a detection plate, wherein the detection plate is coated with duck circovirus Cap protein; wherein the duck circovirus Cap protein is obtained through expression of a eukaryotic expression system. The Cap protein prepared by using an eukaryotic expression system is used as a coating antigen, has good antigenicity, can be specifically combined with a duck circovirus antibody in a serum sample, and ensures that the provided kit has very strong specificity and sensitivity to the duck circovirus. According to the duck circovirus ELISA antibody detection kit provided by the invention, the duck circovirus antibody can be rapidly and accurately detected, and the problems of poor antigen quality and unstable detection performance of the existing duck circovirus ELISA antibody detection method are solved.

This invention belongs to the technical field of virus detection, specifically relating to a PCR detection kit for the simultaneous detection of four DNA viruses in waterfowl and its application. The PCR detection primer set for the simultaneous detection of four DNA viruses in waterfowl of this invention is the optimal primer set obtained through experimental screening. The kit composed of these primers can rapidly and specifically amplify and diagnose Muscovy duck parvovirus, duck plague virus, duck adenovirus type 3, and duck circovirus simultaneously, exhibiting high specificity and sensitivity. It can accurately and rapidly identify MDPV, DPV, DAdV-3, and DuCV, and can complete quantitative real-time PCR detection in approximately 50 minutes. The kit is simple and practical to operate, achieving efficient simultaneous detection of four viruses, shortening the detection cycle, increasing sensitivity by 10-1000 times, significantly improving early disease diagnosis capabilities, significantly reducing the misdiagnosis rate of diseases in poultry farms, and contributing to the sustainable development of the poultry farming industry.

The invention discloses a method for separating and amplifying duck circovirus by using duck embryo primary hepatocytes, and relates to the technical field of animal virology and cell culture. According to the method, a trypsin-EDTA (Ethylene Diamine Tetraacetic Acid) and collagenase IV double-enzyme digestive system is creatively combined and the HGF is added, so that the yield and the activity of primary hepatocytes are remarkably improved, and the duck circovirus is quickly separated on the basis of the yield and the activity of the primary hepatocytes. According to the technical scheme, the duck embryo liver is adopted as a cell source, the cell survival rate is increased by 16.5% by shortening the digestion time and optimizing the enzyme ratio, an HGF stable cell membrane structure is innovatively introduced, a complete DuCV in-vitro proliferation system is established, and the virus titer reaches a peak value 60 hours after inoculation. According to the method, the duck circovirus is rapidly separated and amplified by using the duck embryo primary hepatocytes, and the living cell percentage of the primary cells is improved, so that the problems of low duck circovirus separation efficiency, low primary cell utilization rate, high cost and the like in the existing method are solved.

The present application relates to poultry disease immune control technology, and in particular to a duck circovirus genetic engineering subunit vaccine and a preparation method and application thereof. A primer group shown as SEQ ID NO. 1 and SEQ ID NO. 2 is used to perform PCR amplification on a target Cap gene with a nuclear localization signal removed, a prokaryotic expression vector pET28-cap containing a His tag is constructed, is transferred into a BL21 (DE3) competent cell for induction expression, and the cap protein is purified by using a NI purification resin. After denatured cap protein after purification is renatured, the antigen can be obtained, is emulsified with white oil adjuvant to prepare a vaccine. The present application successfully realizes prokaryotic expression of the cap protein by designing primers of the Cap gene with the nuclear localization signal removed, and shows good immunogenicity, and can be used for development of duck circovirus genetic engineering subunit vaccine, egg yolk antibody and the like.

The application belongs to the field of genetic engineering and relates to an shRNA for inhibiting duck circovirus gene expression, wherein the shRNA is artificially synthesized, can inhibit duck circovirus Rep and Cap gene expression, and thus plays a role in inhibiting virus replication in a duck body. Experiments prove that injection of recombinant plasmids containing the shRNA interference sequence can significantly inhibit virus replication in a duck body, and compared with a control group, the virus copy number is reduced by 100 times at most, which has important significance in clinical prevention and control of duck circovirus.