9 results about "Duck circovirus" patented technology

The invention belongs to the technical field of molecular biology, and particularly relates to a dual fluorescent quantitative PCR (polymerase chain reaction) primer probe group for detecting goose circovirus and duck circovirus type II and application of the dual fluorescent quantitative PCR primer probe group. The fluorescent quantitative PCR primer probe group comprises a primer pair and a probe, and the sequences of the primer pair and the probe are as shown in SEQ ID NO.1 to SEQ ID NO.6. The fluorescent quantitative PCR primer probe group has good specificity on the goose circovirus and the duck circovirus type II, the lowest detection amount of GoCV detection is 2.46 * 10 < 2 > copies / mu L, and the lowest detection amount of DuCV-II detection is 2.5 * 10 < 2 > copies / mu L. Besides, in-batch and inter-batch real-time fluorescent quantitative PCR amplification is carried out on a standard substance, the in-batch variable coefficient is less than 1%, and the inter-batch variable coefficient is less than 1.6%, so that the primer probe group and the kit provided by the invention are high in stability and good in repeatability.

The invention relates to a dual LAMP (loop-mediated isothermal amplification) detection system as well as a kit and a detection method thereof. The dual LAMP detection system comprises a primer group, wherein the primer group comprises four primers for specifically detecting the cap gene of the duck circovirus and four primers for specifically detecting the Fiber2 gene of the duck adenovirus type 3. The invention provides a duck circovirus and duck adenovirus type 3 dual LAMP detection system which is efficient, accurate and easy to operate and a corresponding kit. The kit does not need to be operated by complex equipment or professionals, can detect the duck circovirus and the duck adenovirus type 3 at the same time, can introduce a visual detection technology, is suitable for grassroots veterinary laboratory and on-site rapid detection, provides a new detection method for the duck circovirus and the duck adenovirus type 3 for grassroots veterinarians and farmers, and can be used for detecting the duck circovirus and the duck adenovirus type 3 at the same time. Therefore, the requirement on rapid diagnosis of the two viruses in practical application is met.

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.

The invention discloses a separation method for separating duck circovirus by using SPF duck embryos, belongs to the technical field of biological vaccine prevention and treatment, and solves the technical problem that in the prior art, the duck circovirus does not successfully obtain a virus breeding method. The separation method for separating the duck circovirus by using the SPF duck embryo, provided by the invention, comprises a duck circovirus inoculation mode, a virus inoculation method and a method for obtaining a virus culture. The virus separation method is easy and convenient to operate, economical, suitable and small in potential infection risk, and can be used as a common research method for duck circovirus separation, identification, antigen preparation and the like. So far, the method is of great significance to research on the pathogenesis and pathogenesis of the immunosuppressive disease caused by the duck circovirus.

The invention relates to a reagent combination and a kit for visually detecting duck circovirus based on LAMP-CRISPR / Cas12a and application of the reagent combination and the kit, the reagent combination comprises LAMP amplification primers and crRNA, the LAMP amplification primers comprise two outer primers F3-1 and B3-1 and two inner primers FIP-1 and BIP-1, the crRNA is any one of crRNA-1, crRNA-2 and crRNA-3, and the sequences of the LAMP amplification primers and the crRNA are shown as SEQ ID NO.1-4 and SEQ ID NO.13-15. The invention further discloses a kit for visually detecting duck circovirus based on LAMP-CRISPR / Cas12a. According to the method, the LAMP-CRISPR / Cas12a detection system is established by using the reagent combination, so that the rapid detection of the duck circovirus is realized. The detection method established by the invention does not need an expensive thermal cycle instrument, is simple to operate, and can achieve the purpose of rapid visual detection.