57 results about "Goose astrovirus" patented technology

The invention provides a real-time fluorescence quantification PCR detection primer for goose astroviridae and a kit of the detection primer. The primer has the sequences represented by SEQ ID NO.1-2 and has very high specificity and sensitivity. Currently, a primer for a real-time fluorescence quantification PCR detection method for detecting the goose astroviridae based on saturated fluorescent dye Eva Green has not been researched and reported at home and abroad, and blanks in the relevant fields at home and abroad can be filled up by virtue of the detection primer and the kit.

The invention discloses an egg yolk antibody for preventing novel goose astrovirus and a preparation method thereof. The egg yolk antibody contains a goose astrovirus strain resistance antibody; the preservation number of the goose astrovirus strain is CCTCC NO:V201808. The prepared egg yolk antibody is good in safety, and no any partial or whole adverse reaction is caused by the egg yolk antibody; infections caused by novel goose astrovirus can be effectively prevented and / or treated, and the good commercialization development prospect is achieved.

The invention belongs to the field of epizootiology detection, and discloses a goose astrovirus loop-mediated isothermal amplification detection primer group and a kit. According to the method, primers are designed according to goose astrovirus genome conserved sequences: outer primers F3 and B3; inner primers FIP and BIP; a loop primer LB. The specific sequences are shown in SEQ ID NO.1-5 separately. According to the designed primers, a loop-mediated isothermal amplification method for detecting goose astroviruses is established. According to the detection method, no cross reaction with common goose common goose infectious disease pathogens occurs. The detection method is high in specificity, good in accuracy, high in sensitivity, free of expensive instruments and equipment and convenient and facilitates public use.

The invention discloses an inactivated vaccine for preventing and treating novel goose astroviruses and a preparation method thereof. The inactivated vaccine is prepared from the effective dose of inactivated goose astrovirus strain virus liquid in prevention or treatment. A preservation number of the goose astrovirus strain is CCTCC NO: V201808. The prepared novel goose astrovirus inactivated vaccine is good in safety, any local or general adverse effects caused by the vaccine do not appear, and the detection of each index is stable and effective. After the inactivated vaccine is used for immunizing a young goose, the young goose can acquire the higher antibody level, and a persistent period is long. The infection and outbreak of the novel goose astroviruses can be prevented and treated specifically, and the effective immune protection is provided for a goose group. In addition, a production technology for the vaccine is simplified, and the large-scale production can be realized.

The invention discloses a goose astrovirus egg-yolk antibody compound and a preparation method thereof and relates to the field of veterinary biopreparations. To achieve the objective, the preparation method comprises the steps of 1) using a goose astrovirus SD strain collected under CCTCC NO: V201815 as a vaccine producing viral strain to prepare an inactivation antigen; 2) subjecting the inactivation antigen to water phase preparation and oil phase preparation, mixing a water phase and an oil phase, and performing primary emulsification to obtain an immune antigen; 3) inoculating the immune antigen to layers so as to produce high-immunity eggs; and 4) collecting yolk from the high-immunity eggs, disinfecting, separating yolk, diluting the yolk, acidifying, extracting, preparing a stock antibody solution, filtering, concentrating, and inactivating to obtain the goose astrovirus refined egg-yolk antibody. Goose astrovirus can be prevented effectively, geese can be protected within the whole growth period, and farming benefit can be increased evidently.

The invention discloses a novel goose astrovirus which is preserved in the China Center for Type Culture Collection on April 26, 2018, and the preservation number is CCTCC NO: V201808. The novel gooseastrovirus disclosed by the invention has good immunogenicity upon popular goose astroviruses with a main symptom of gosling gout at present. An inactivated vaccine prepared from a novel goose astrovirus strain disclosed by the invention is good in security, has a protection rate up to 100%, and is capable of providing complete protection on newly separated goose astrovius variants.

The invention provides an egg yolk antibody prepared by using new goose astrovirus. The egg yolk antibody is prepared by taking inactivated vaccine prepared by using a new goose astrovirus GD 0122 strain with the preservation number of CCTCC No:V201820 as an antigen. The prepared antibody is good in safety, after a gosling is inoculated with the antibody, no local or systemic adverse reactions caused by the injection of the antibody occur; the prepared antibody is evaluated by adopting an antibody titer determination and challenge protection test, the result shows that the agar diffusion antibody titer of the prepared antibody is not lower than 1:16, after the antibody is injected, the gosling can resist the attack of virulent virus, and the prepared antibody can provide effective immune protection for geese and has a good commercial development prospect.

The invention discloses a fluorogenic quantitative PCR kit for detecting novel astrovirus of geese and application of the kit. The fluorogenic quantitative PCR kit is internally provided with a primershown as SEG ID NO.1-SEQ ID NO.2, a probe shown as SEQ ID NO.3, a standard substance and a fluorogenic quantitative PCR reaction reagent. The fluorogenic quantitative PCR kit can achieve detection onthe novel astrovirus of geese, is excellent in specificity and can only be specifically combined with the novel astrovirus of geese without any cross reaction with other goose source viruses; the detection sensitivity is high, a great linear relation can be maintained within a (4*100)-(4*109) copy range of the standard substance, the detection range can reach 10 orders of magnitude, sick goose tissue of geese carrying the novel astrovirus and virus carrier geese without clinical symptoms can be detected efficiently, and the detection efficiency is improved.

The invention discloses a novel goose astrovirus SYBR Green dye method fluorescent quantitative PCR detection kit. The fluorescent quantitative PCR detection kit contains primer pairs shown in SEQ IDNO.1-SEQ ID NO.2, standard plasmids and fluorescent quantitative PCR reaction reagents. The fluorescent quantitative PCR kit can detect a novel goose astrovirus, the kit is high in specificity, is only combined with the specificity of the novel goose astrovirus, and has no cross reaction with other goose-origin viruses; the detection sensitive is high, the quite good linear relation is achieved within the 1*10<1>-1*10<7> coping range of standards; novel goose astrovirus diseased goose tissue and virus carrier gooses with no clinic symptom can be effectively detected, and the detection efficiency is improved.

The invention discloses a preparation method and a product of a goose astrovirus egg yolk antibody. The preparation method comprises the following steps: (1) preparing an astrovirus liquid and an adjuvant, using the astrovirus liquid and the adjuvant as immunogens to immunize a healthy laying goose to prepare a highly immunized egg, collecting the highly immunized egg; (2) disinfecting the highlyimmunized egg, collecting an egg yolk; and extracting the egg yolk antibody from the egg yolk by using an acid water-carrageenan method. The recovery rate of the goose astrovirus egg yolk antibody extracted through the preparation method is more than 93%, the purity is more than 95%, and the indexes like the recovery rate, the extracting quantity and the purity of the goose astrovirus egg yolk antibody are obviously better than those of the goose astrovirus egg yolk antibody prepared by using the existing method. The goose astrovirus egg yolk antibody prepared by using the preparation method achieves the better protecting effect only by 0.5 ml of a preventive dose, and the rate of protection against the virulent strain reaches 100%. The infection and the outbreak of the goose astrovirus isprevented specifically, meanwhile, the production process is simplified, and the preparation method can implement large-scale production and is applied clinically.

The invention provides a novel goose-star virus which can cause gosling gout and is provided with a GD 0122 strain. The strain is preserved in a Typical Chinese Culture Preservation Center of the Wuhan university on May 13, 2018, and the collection number of the strain is CCTCC No:V201820. The novel goose-star virus can be used for preparation of vaccines or antibodies for preventing gosling gout.The gosling gout vaccines prepared by the selected virus is good in safety, and any local and whole body adverse reaction caused by the vaccines are omitted. In a storage life test, character, safetytest and potency test data are analyzed, various indexes are stable and effective, immune effects of the vaccines are assessed by a serological method and an immune attacking method, results show that the prepared inactivated vaccine can provide effective immune protection for geese, and the virus has an excellent commercial development prospect.

The invention provides a preparation method of attenuated goose astrovirus and a live vaccine prepared by applying the attenuated goose astrovirus, and belongs to the technical field of waterfowl disease prevention and control. A method combining high-temperature treatment and low-temperature incubation is adopted to attenuate the virus. The method has the advantages of short virus attenuation time, low cost and long immune protection time. There is virulence return after multiple passages, thus effectively preventing goose gout disease caused by goose astrovirus.

The invention relates to the technical field of virus detection, in particular to a multiple fluorescent quantitative PCR detection primer pair for novel goose astrovirus, goose paramyxovirus and goose parvovirus, and an application of a specific probe. The established multiple fluorescent quantitative PCR system can accurately detect single or mixed infection of the novel goose astrovirus, the goose paramyxovirus and the goose parvovirus, the multiple fluorescent quantitative PCR system is optimized, the optimal primer concentration and the optimal probe concentration are determined, the detection precision is higher, and the multiple fluorescent quantitative PCR detection primer pair has important advantages in the field of virus identification.

The invention discloses an immune composition which comprises goose astrovirus outer capsid structure protein coded by adopting nucleic acid molecules with a sequence shown as SEQ ID NO:1 or higher than 95% identical nucleic acid molecules of a nucleotide sequence shown as SEQ ID NO:1. The immune composition can be applied as a goose astrovirus-like particle novel genetic engineering subunit vaccine, is high in antigen expression level, can be spontaneously assembled into virus-like particles (VLPs) and is high in immunogenicity and apathogenic to gooses. The vaccine can be prepared on a largescale by using a bioreactor through serum-free suspended culture, and vaccine production cost is lowered greatly.

The invention relates to an ELISA kit used for detecting goose astrovirus antibodies, and a preparation method thereof. The ELISA kit used for detecting goose astrovirus antibodies comprises an enzymelabeled plate, an enzyme labeled second antibody, goose astrovirus positive reference serum, goose astrovirus negative reference serum, a diluents, 10* washing liquid, TMB substrate solution, and a stopping solution. The antigen used for coating the enzyme labeled plate is goose astrovirus Cap protein; the goose astrovirus is Avastrovirus III type. According to the ELISA kit, expression purifiedgoose astrovirus (Avastrovirus III type) Cap protein is taken as the coating antigen; indirect ELISA method is adopted in detection of goose serum samples to be detected; and the kit used for detecting goose astrovirus antibodies in serum is assembled. The ELISA kit can be used for goose astrovirus infection serology diagnosis and antibody level monitoring, and epidemiological investigation, and asimple and rapid method is provided for goose astrovirus prevention and treatment.

The invention relates to the technical field of virus detection, and specifically relates to multiplex PCR detection primer pairs for goose astrovirus, goose paramyxovirus and goose parvovirus; and the invention further relates to a detection method of the primer pairs and application of the primer pairs. The multiplex PCR system established by the invention is capable of accurately detecting simplex or mixed infection of waterfowl with the goose astrovirus, the goose paramyxovirus and the goose parvovirus. Moreover, the multiplex PCR system is optimized with determined optimal primer ratio, primer concentration and Premix rTaq enzyme; and thus, better DNA amplification effects of the 3 viruses are achieved. Therefore, the multiplex PCR system has higher detection accuracy, and thereby having important advantages in the field of virus identification.

The invention discloses a novel goose astrovirus with cross-species transmission capacity and application thereof, the virus is preserved in China Center for Type Culture Collection on January 12, 2020, and the preservation number of the virus is CCTCC NO: V202019. The novel goose astrovirus provided by the invention has very good immunogenicity on currently popular goose astroviruses taking duckling nephritis as a main disease. An inactivated vaccine prepared from the novel goose astrovirus strain is good in safety, the protection rate reaches 100%, and complete protection can be provided fornewly separated goose astrovirus variants.

The invention provides a goose astrovirus variation strain. The virus is a variation strain of classical goose astrovirus FLX, and the preservation number is CCTCC NO: V201976. The screened virus strain is applied to preparation of vaccine. Isolation, identification and purification of pathogens is performed on clinical gosling gout samples so as to obtain the goose astrovirus strain, and it is confirmed through observation of a transmission electron microscope, RT-PCR tests, serological tests and animal regression tests that the screened virus is the variation strain of the classic goose astrovirus FLX; and the variation strain has significant differences of serological and biological features from the classic strain, and a basis is provided for comprehensive prevention and control of thedisease in goose farms in China.

Provided is a preparation method of a goose gout yolk antibody. A goose astrovirus RPVA1061 strain is separated from pathological livers and kidneys of clinically infected goslings, and the RPVA1061 strain is used for preparing the yolk antibody for prevention and treatment of clinical gosling gout; the problem of no medicine for a conventional clinical goose gout is solved, and a new clinical prevention and control scheme of goose gout is provided; through clinical verification, the prepared yolk antibody has remarkable prevention and control effects on goose gout.

The invention relates to a method for efficiently expressing an ORF2 (Open Reading Frame 2) gene of a goose astrovirus soluble capsid precursor, and application of the method, and belongs to the fieldof veterinary biological products. The ORF2 gene of the goose astrovirus is divided into two-segment expression according to the difference of gene sequence functions, and expression products are independently capsid precursor (short for cap) and capsid spike (short for spike). Two segments of proteins segmented according to functional zones can be expressed by escherichia coli, baculovirus and aCHO (Chinese Hamster Ovary) expression system. Through a purification method, the capsid precursor expression protein and the capsid spike expression protein of the goose astrovirus can be obtained.The expressed capsid precursor and capsid spike can be used for preparing a subunit vaccine and an egg yolk antibody of the goose astrovirus. The subunit vaccine and the egg yolk antibody prepared bythe method disclosed by the invention have a good immune effect and treatment effect, can be used for preventing diseases caused by the goose astrovirus and have the advantages of being in low in production cost, simple in operation and good in biological safety.

The invention discloses an egg yolk antibody for preventing and treating a novel goose astrovirus with cross-species transmission capability. Inactivated novel goose astrovirus with a preservation number of CCTCC NO: V202019 is used as an antigen to prepare vaccine immune laying hens, and the egg yolk antibody is extracted and purified from high-immunity egg yolk. The goose astrovirus with the preservation number of CCTCC NO: V202019 is used as a vaccine production strain to prepare an inactivated vaccine to immunize laying hens, the egg yolk antibody can be efficiently obtained, the egg yolkantibody is good in safety, and any local or whole-body adverse reaction caused by the egg yolk antibody does not occur; and moreover, the egg yolk antibody can prevent and treat infection of the novel goose astrovirus effectively and has good commercial development prospects.

The invention discloses a goose astrovirus bivalent inactivated vaccine, an egg yolk antibody and preparation methods thereof. According to the goose astrovirus bivalent inactivated vaccine, acclimatized LMH cells are inoculated with goose astrovirus type 1 strains and goose astrovirus type 2 strains separately for suspension culture, and a virus solution which is high in virus content and free ofconcentration is obtained; the goose astrovirus bivalent inactivated vaccine is obtained by mixing and emulsifying inactivated virus solution and a vaccine adjuvant; the goose astrovirus bivalent eggyolk antibody is obtained by immunize laying hens with the inactivated vaccine to obtain high-immunity eggs and then separating the egg yolk and extracting antibody. The goose astrovirus bivalent inactivated vaccine and the egg yolk antibody prepared by the invention can provide effective immune protection for goslings, can effectively prevent goose gout, and have a good commercial development prospect.

The invention discloses an indirect ELISA detection kit for detecting a novel goose astrovirus antibody. The kit includes an ELISA plate coated with a novel goose astrovirus ORF1b protein recombinantantigen. The preparation method of the novel goose astrovirus ORF1b protein recombinant antigen includes: taking the complete gene sequence of goose astrovirus with a preservation number of CCTCC NO:V201808 as the template, conducting primer pair amplification to obtain an ORF1b gene fragment, constructing a recombinant expression vector, and expressing the goose astrovirus ORF1b protein recombinant antigen by a prokaryotic expression system. The indirect ELISA detection kit provided by the invention can achieve rapid and effective detection of the new outbreak novel goose astrovirus antibodywith gout as the main symptom, thus monitoring the epidemic situation of novel goose astrovirus in a gaggle of geese.

The invention relates to a goose astrovirus (GoAstV), goose parvovirus (GPV) and goose calico virus (GoCV) multiple nano PCR detection primer pair, a kit and an application method thereof. The kit comprises a reverse transcription mixed solution, an amplification reaction mixed solution, a negative control and a positive control. According to the method, a nano PCR detection application method for the goose astrovirus, the goose parvovirus and the goose calico virus is established, a multiple nano PCR reaction system is optimized, and the primer concentration, the annealing temperature, the rTaq DNA polymerase concentration, the nano particle diameter and the nano particle concentration are determined, so that the amplification effects of the three viruses of GoAstV, GPV and GoCV are better; and the method is high in sensitivity, good in specificity and high in stability, the detection method is easy to operate, convenient and rapid, the detection time can be greatly shortened, the diagnosis problem of GoAstV, GPV and GoCV mixed infection can be thoroughly solved, and the method is suitable for rapid detection in clinical laboratories.

The invention discloses real-time fluorescence quantification PCR (polymerase chain reaction) detection primers for a novel goose arbovirus which causes gout of goslings, and a kit of the primers. Thekit comprises an upstream primer GoAstV-F1 and a downstream primer GoAstV-R1. The kit disclosed by the invention is capable of detecting the novel goose arbovirus which causes gout of the goslings, has the advantages of being good in specificity, good in repeatability and accurate in quantification, is good in sensitivity, is capable of detecting 6.8 copies at least, has a very good linear relationship within a 6.88-6.88*10<9> copy range of a standard product, has a detection range up to 10 orders of magnitudes, is capable of rapidly detecting the novel goose arbovirus GoAstV which causes gout of the goslings, and improves the detection efficiency.

The invention relates to an ELISA kit for detecting goose astrovirus antibody and a preparation method thereof, comprising an enzyme-labeled plate, an enzyme-labeled secondary antibody, a goose astrovirus positive control serum, a goose astrovirus negative control serum, a diluent, and a 10× Washing solution, TMB substrate solution, and stop solution; the antigen coated on the microtiter plate in the kit is the Cap protein of goose Astrovirus; the goose Astrovirus is avian Astrovirus type 3. The present invention expresses and purifies newly occurring Goose Astrovirus (Avian Astrovirus Type 3) Cap protein as the coating antigen, uses the indirect ELISA method to detect the goose serum sample to be tested, and assembles it for the determination of the Astrovirus antibody in the serum kit. The kit can be used for serological diagnosis of recent goose astrovirus infection, monitoring of antibody levels, epidemiological investigation, etc., and provides a simple and quick method for the prevention and control of goose astrovirus.

The invention discloses an ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit for a goose astrovirus Capsid protein antigen, a detection method and application. The kit comprises: an ELISA platecoated with a goose astrovirus Capsid protein polyclonal antibody, a confining liquid, a sample diluent, a purified goose astrovirus Capsid recombinant protein antigen standard substance, an HRP labeled anti-goose astrovirus Capsid monoclonal antibody, a washing liquid, an enzyme substrate solution A, an enzyme substrate solution B and a stop solution; meanwhile, the invention further discloses amethod for accurately and quantitatively detecting the goose astrovirus Capsid protein antigen in a sample by adopting the kit. The kit contains an antigen standard substance, can detect the accuratecontent of the goose astrovirus Capsid protein in a sample, and is suitable for quantitative detection of the goose astrovirus Capsid gene subunit vaccine.

The invention relates to a primer pair, a kit and a method for detecting goose astrovirus and goose calicovirus. The primer pair disclosed by the invention comprises a GoAstV upstream primer, a GoAstV downstream primer, a GoCV upstream primer and a GoCV downstream primer which are respectively shown as SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4. The kit disclosed by the invention comprises the primer pair. The method comprises the following steps: S1, extracting RNA (Ribonucleic Acid) of a sample; S2, adopting the primer pair, amplifying by using a dual one-step RT-PCR reaction program, and then detecting; if a strip appears at 452 bp, indicating that the goose astrovirus is positive, and if a strip appears at 711 bp, indicating the goose calico virus is positive. The scheme provided by the invention is used for molecular epidemiological detection of goose astrovirus and goose calico virus, and is good in specificity and reliable in result.

The invention relates to a goose astrovirus N-GoAstV-190123 with strong virulence and good immune performance, after the goose astrovirus N-GoAstV-190123 is prepared into an inactivated vaccine for immunization, a relatively high neutralizing antibody can be generated, which can be used to protect a goose group completely, and has an important economic value for the goose breeding industry.

The invention belongs to the technical field of virus detection, and particularly relates to a novel goose astrovirus (GoAstV) and duck Tembusu virus (DTMUV) multiplex fluorescent quantitative PCR detection primer probe group, a kit and application. According to the application, specific primers and probes are designed respectively aiming at an E gene of DTMUV and an ORF1b gene of GoAstV, and a fluorescent quantitative PCR method for simultaneously detecting DTMUV and GoAstV in the same system is established by optimizing reaction conditions. The method has the advantages of specificity, sensitivity and high efficiency.