55 results about "Avian leukosis" patented technology

The invention belongs to the technical field of animal epidemic disease serological diagnosis, and relates to an enzyme-linked immunosorbent assay vector and a kit for detecting avian leukosis P27. The avian leukemia virus (P27) enzyme linked immunosorbent assay kit comprises a 96 hole elisa plate coating a avian leukosis P27 polyclonal antibody, P27 monoclonal antibody marked by alkaline phosphatase, a substrate solution, a stop solution and a cleaning solution. The vian leukosis P27 polyclonal antibody and the P27 monoclonal antibody marked by alkaline phosphatase effectively improve the sensitivity, the specificity and the stability of the detection. The invention provides an efficient and sensitive ELISA (enzyme linked immunosorbent assay) detection kit for sifting out and purifying avian leukosis positive chickens and cultivating new species with heredity resistance, and the kit is low in cost and simple and convenient to operate, and is applicable to promotion in animal husbandry.

The invention provides a fowl leucovirus subgroup J specific antigenic polypeptide and an application thereof. The sequences of antigenic polypeptide are SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 or deriving sequences of the sequences above. Besides, the antigenic polypeptide is coded by genes in gp 85 section of envelope protein gene (env) of a virus, and has strong specificity, good hydrophily, good reactivity and high purity. In addition, the antigenic polypeptide is used for preparing an indirect ELISA detection kit which can conduct specific detection on an ALV-J antibody, wherein the detection kit comprises one or more of enzyme labeled plates of ALV-J specific antigenic polypeptide above and related reagents, and the enzyme labeled plates and the reagents coat the detection kit. During the detection of ALV-J antibody, the kit has high specificity and low cost, is beneficial to popularization, and the effects are obviously superior to those of conventional kits and imported kits. The ALV-J specific antigenic polypeptide and the ELISA detection kit prepared by the antigenic polypeptide are applicable to evaluation of results of infection or purification of the ALV-J antibody; the ALV-J specific antigenic polypeptide is applicable to preparation of antibody of specific ALV-J.

The invention discloses an LAMP (Loop-Mediated Isothermal Amplification) kit for detecting a main subtype avian leukemia virus. A loop-mediated isothermal amplification (LAMP) technology is adopted, and two pairs of specific primers (inner primers FIP and BIP, outer primers F3 and B3) are designed according to a POL (Point Of Load) gene sequence of the avian leukemia virus. By applying the LAMP kit and a detecting method established by the LAMP kit, an LAMP real-time turbidity meter is utilized to carry out real-time, quantitative and whole-course sealed monitoring analysis on LAMP reaction primers of the ALV (Avian Leukemia Virus), a reaction system and a reaction process, so that LAMP primers of the ALV are efficiently and specifically amplified. The LAMP kit disclosed by the invention has the advantages that the specificity is strong, the sensitivity is high, a result is quickly obtained, pollution is not caused and a product can be detected in real time; the avian leukemia virus can be detected out in real time by sampling according to the established system, and the detected result can be quickly and accurately obtained, so that convenience is brought for simply and quickly detecting the avian leukemia virus.

The present invention relates to prokaryotic expression of fowl leukemia related p27 protein and its application. The present invention constitutes new recombinant expression plasmid with the designed specific primer, so as to express high level and high purity p27 protein successfully in prokaryotic cell.

The invention discloses a chicken B,D,E-subgroup avian leukaemia genetic resistance related mononucleotide polymorphism molecular marker and application thereof. The molecular marker is TVB gene DNA (deoxyribonucleic acid) sequence SNP-3674C/T, or TVB gene coding sequence SNP-298C/T. The molecular marker can be used for identifying chicken B,D,E-subgroup avian leukaemia genetic resistance characteristics; correspondingly, the molecular marker can be distinguished by a PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) process, and can be used for resistance breeding to control the risk of B,D,E-subgroup avian leukaemia from the source; and the molecular marker has the advantages of high purposiveness and the like, is simple to operate and easy to implement, greatly lowers the screening cost, and thus, has important practical meanings.

The invention discloses a livestock seminal fluid disinfectant, a preparation method thereof and an application. The disinfectant consists of the following components by weight percent: 1.0 to 2.0 percent of radix isatidis extract, 1.0 to 2.0 percent of melia azedarach extract, 1.0 to 2.0 percent of sanguisorba officinalis extract, 1.0 to 2.0 percent of folium isatidis extract, 1.0 to 2.0 percent of aloe extract, 2.0 to 3.0 percent of glucose/ fructose, 3.0 to 5.0 percent of glycerinum, 2.0 to 3.0 percent of sodium citrate, 0.2 to 0.3 percent of vitamin, and the balance of distilled water, wherein the vitamin is one or a combination of more than two of vitamin C, vitamin A and vitamin E. The disinfectant can inhibit or kill hog cholera virus, porcine circovirus, avian leukosis and pullorum disease salmonella pathogen. A main formula of the seminal fluid disinfectant is a pure traditional Chinese herba preparation, so that the disinfectant has low toxicity, no residue and good safety; and by virtue of the test for the negative influence on the activity of sperms, the disinfectant is small in toxicity and is safe and reliable.

The invention discloses a loop-mediated isothermal amplification reaction primer for detecting B substock avian leukosis, which is composed of a pair of outer primers, a pair of inner primers and a pair of loop primers, wherein the outer primers are composed of a primer with SEQ ID NO.1 and a primer with SEQ ID NO.2; the inner primers are composed of a primer with SEQ NO.3 and a primer with SEQ NO.4; and the loop primers are composed of a primer with SEQ ID NO.5 and a primer with SEQ ID NO.6. In the loop-mediated isothermal amplification reaction primer for detecting the B substock avian leukosis provided by the invention, a B substock LAMP primer is designed by selecting an avian leukoviruses gp85 section, and according to the designed primer, a method for detecting ALV-B type LAMP is built; the detection method can not be used for carrying out cross reaction with other main substocks of the avian leukosis and can not be used for carrying out heterogenetic reaction with other common poultry communicable diseases at the same time; and the detection method is convenient and accurate, the local applicability is good, the accuracy is high, the specificity is strong and the sensitivity is high.

RT-PCR based systems for detection of exogenous and endogenous avian leukosis/sarcoma viruses in the albumen of fertilized and unfertilized poultry eggs have been developed. Such systems permit the identification of virus-contaminated eggs and of viral presence in sera of birds. These systems aid in the prevention of high loss rates of egg-laying birds in the poultry industry and the reduction in potential human exposure to these oncogenic retroviruses through consumption of contaminated commercial eggs. It is further envisioned that detection methods can be modified to include viral detection of other samples from various avian origins and other animal or human retroviruses.

The invention discloses a method for reducing a chicken breeding flock avian leukosis positive rate. Under the condition that the number of negative cocks does not reach a breed conservation requirement, according to a method for processing seminal fluid of positive cocks, under the condition that sperm motility is not influenced, the content of viruses in the seminal fluid of the positive cocks is reduced as far as possible, and then breed mating is carried out on the cocks and the negative hens, namely continuity of flocks is ensured, the positive rate of chicken flocks of a next generation is also effectively reduced, and finally negative core flocks are established after replacement of a few generations.

The invention belongs to the technical field of poultry breeding, and discloses a chicken avian leukosis purification method. The method comprises the following steps: a safety infrastructure is established, and safety management is implemented in a breeding process; in the safe breeding process, leukemia purification is carried out on the chickens; in the avian leukosis purification process, different detection indexes are collected according to different breeding stages, and positive chickens are removed; and the infrastructure is cleaned and disinfected on time in the breeding process, the negative chickens are put into the infrastructure for breeding expansion, and the negative chickens are injected with vaccines. When the negative chickens are placed in the infrastructure for breeding expansion, the negative chickens are used as breeding for isolated feeding, and the negative chickens are placed in the same cage; and a blocking measure is adopted between the cages, so that horizontal propagation is avoided, and direct contact between the cages and direct airflow convection are avoided. Meanwhile, the problem that purification of avian leukosis is divided into three stages of reduction, control and eradication in the prior art, and the difficulty is sequentially increased is solved.

The invention belongs to the technical field of animal disease detection, and discloses a specific antigen epitope for avian leukosis virus subgroup J, a fusion protein, a specific antibody and an application thereof. The amino acid sequence of the specific antigen epitope for avian leukosis virus subgroup J is shown in SEQ ID NO: 1. Experiments show that the antigen epitope responses to the serum of the subgroup J and does not responses to the serum of the subgroup AB, and can specifically recognize the avian leukosis virus subgroup J. The fusion protein can be prepared into a vaccine to stimulate a receptor to generate an antibody and protect the receptor against the infection of the avian leukosis virus subgroup J. The fusion protein can immunize animal to prepare the specific antibody of the avian leukosis virus subgroup J. The specific antibody of the avian leukosis virus subgroup J can be specifically recognized with the specific antigen epitope for avian leukosis virus subgroup J, and a detection kit is developed to distinguish the avian leukosis subgroup AB and the avian leukosis subgroup J.

The invention provides a fowl leukemia purification method, comprising the steps of determining a breeding target, screening individuals that basically meets the characteristics of the breeding target, numbering all individuals to form an F0 generation breeding population, adopting single-cage feeding, conducting artificial insemination; adopting an ALV P27 antigen detection method to conduct breeding screening on fowls from F0 to F4; and finally conducting 4-5 generations of purification through a breeding core group until ALV P27 antigen detection reaches zero detection, so as to realize fowl leukemia-negative specialized strain cultivation. According to the invention, breeding work and disease purification work are organically combined to form a fowl leukemia breeding purifying group, and a specialized fowl leukemia-negative strain is cultivated for commercial matching production. The method of the invention completely solves the purification problem of fowl leukemia from the provenance.

The invention discloses a PCR detection primer group of avianleukosis (AL) virus and a kit comprising the detection primer group. The PCR detection primer group of the avianleukosis virus comprises primers P1, P2, P3 and P4, wherein P1 is an upstream universal detection primer; P2 is a downstream primer for detecting ALV-A subtype virus; P3 is a downstream primer for detecting ALV-B subtype virus; and P4 is a downstream primer for detecting ALV-J subtype virus; and the diagnosis kit comprises the PCR detection primer group of the avianleukosis virus, ALV-A subtype virus positive control plasmid, ALV-B subtype virus positive control plasmid, ALV-J subtype virus positive control plasmid and negative control. With the application of the PCR detection primer group of the avianleukosis virus, it can rapidly and accurately detect whether poultry is affected with the leukemia subtype virus or not.

The invention belongs to the technical field of resistant variety breeding, and in particular discloses a chicken B subgroup avian leukosis resistantmolecular marker tvb<3731-3732insA> and a molecular diagnosis method thereof. The invention analyzes genetic variation of a Chinese chicken breed tvb receptor gene for the first time and discovers that the 3731st and the 3732nd sites of the Chinese chicken breed tvb receptor gene have A insertion mutation, and the inventor proves, from in vivo and in vitro experiments two aspects, that the natural mutation causes a host to generate genetic resistance to infection of ALV-B; moreover, the inventor establishes a molecular diagnosis method for a B subgroup avian leukosis resistant genetic marker, and the method can be applied to screening breeding materials of ALV-B genetic resistant chicken varieties (strains) so as to carry out breeding of the ALV-B genetic resistant chicken varieties (strains).

The invention provides a PEG (Polyethylene Glycol) modification based subgroup J avian leukemia virus (ALV-J) immunosuppressive polypeptide and provides a N-terminal alpha-NH2 PEG modification based immunosuppressive polypeptide (ISU) and a preparation method therefor. Compared with the ISU in the prior art, the PEG modified ISU provided by the invention has the advantages that the duplication of ALV-J can be more remarkably suppressed, a scientific basis and a technical support are provided for the prevention and cure of subgroup J avian leukemia and the preparation of PEG modified polypeptide vaccines, and the economic loss of poultry farming caused by ALV-J infection is effectively reduced.

The invention belongs to the technical field of biological products for animals and particularly relates to a vaccine composition and a preparation method thereof. The vaccine composition comprises avian influenza virus H9 subtype antigens, aviadenovirus 4-type antigens, avian reovirus 2-type virus antigens, avian leukosis J-type antigens and an adjuvant. The vaccine composition has good immune effect for the avian influenza virus H9 subtype antigens, the aviadenovirus 4-type antigens, the avian reovirus 2-type virus antigens and the avian leukosis J-type antigens.

The invention relates to an avian leukosis purification field area. The avian leukosis purification field area comprises a life management area, a production area and a dirt treatment area which are isolated by adopting enclosure walls, wherein sewage generated by the purification field area is discharged to a sewage treatment place through a sewage pipe; a vehicle disinfection tank and a personnel disinfection channel are arranged at an entrance of the life management area; a personnel disinfection channel and a shower room are arranged at an entrance of the production area. The invention further relates to an avian leukosis purification method. The avian leukosis purification method comprises requirements of infrastructure in the purification field area, and the requirements of biological safety, production management and purification detection are formulated. The avian leukosis purification field area is compact in layout and reasonable in design, and purification matching measuresare specified in detail, so that the avian leukosis purification field area has practical guiding significance and good operability; according to a purification detection scheme involved in the purification method, a production stage can be flexibly selected for detection according to actual conditions in the field, and detection time points are all avian leukosis detoxification peak periods of breeding hens, so that the infection positive rate of groups is rapidly reduced, and the production performance of purification groups is effectively improved.

The invention belongs to the field of animal husbandry and veterinary and in particular relates to a method for simultaneously purifying and detecting avian leukosis and avian salmonellosis by utilizing one hatching egg and application of the method. The method comprises the following steps: feeding breeder flocks in single cages and encoding; collecting primary hatching eggs of the breeder flocks; enabling the primary hatching eggs to correspond to numbers of the hatching eggs one by one; disinfecting the surfaces of the hatching eggs and opening egg shells from hatching egg air chamber ends;sucking egg white and yolks and putting into PE (Polyethylene) tubes respectively and marking; putting egg white and yolk samples into a refrigerator and freezing and thawing for a plurality of times; taking the frozen and thawed egg white; detecting an ALV virus antigen through an avian leukosis ELISA (Enzyme Linked Immunosorbent Assay) detection method; taking the frozen and thawed yolks and adding normal saline and chloroform; after uniformly mixing on an oscillator, centrifuging and layering a solution, wherein a lower layer is an organic solvent, a middle layer is denatured lipid proteinand an upper layer is an extracted antibody; and sucking the antibody of the upper layer and detecting a salmonellosis antibody through a plate agglutination method; and eliminating chicken flock individuals with positive detection results of the ALV virus antigen and the salmonellosis antibody.

The invention discloses a SNP locus of a chicken NRAMP1 gene. The SNP locus is related to J-subtype avian leukosis resistance, and is nucleotide of the 125 locus located on a NRAMP1 gene segment, the nucleotide of the SNP locus is C or T, and the nucleotide sequence of the NRAMP1 gene segment is shown as SEQ ID NO:1. According to the chicken-NRAMP1-gene SNP locus related to the J-subtype avian leukosis resistance, a specific primer-amplification gene segment containing the SNP locus is designed according to the SNP locus of the 125 locus of the NRAMP1 gene segment, then enzyme digestion analysis is carried out with restriction enzyme, and the polymorphism of mononucleotide cam be simply, rapidly and accurately detected in a low-cost mode. The SNP locus of the chicken NRAMP1 gene is of great significance in the effect mechanism of researching chicken-NRAMP1-gene-resistant J-subtype avian leukosis, purposeful selection on chicken individuals is facilitated, and the resistance of J-ALV to offspring chickens is improved.

The present invention relates to a primer set and a kit for detecting avian leukemia virus A/B/J/K subgroup through one-step PCR, wherein the kit comprises the primer set for detecting avian leukemiavirus A/B/J/K subgroup, ALV-A, ALV-B, ALV-J and ALV-K subtype virus positive control plasmids prepared from the nucleotide sequences amplified with the primer set, and negative control, can rapidly, specifically, sensitively and efficiently detect avian leukemia and distinguish avian leukemia A/B/J/K subgroup, is suitable for epidemiological investigation of avian leukemia, and is contribute to the purification of avian leukemia.

The invention discloses a prevention and purification feeding method of subgroup J avian leukosis of Wuhua Sanhuang chicken. The prevention and purification feeding method comprises the following steps: daily prevention, subgroup J avian leukosis discovery and purification feeding after the discovery of a lesion; 1, daily prevention: uniformly arranging fluorescent lamps in a henhouse and ensuring that the total exposure dosage is enabled to be 9-12 KJ; secondly, mixing a perylenequinonoid compound into feed for feeding from the birth of the chicken; finally, displaying the temperature, the humidity, the luminosity, the chicken quantity and the chicken feeding tracking in the henhouse by using a display device in the henhouse; 2, subgroup J avian leukosis discovery; 3, subgroup J avian leukosis purification treatment: after the discovery of the subgroup J avian leukosis, the treatment dosage is the weight of 40-300 [mu]mol of perylenequinonoid compound/kg per day, the treatment course is 2-4 days, the symptoms of the chicken are observed, and after being normal and being fed for 5-8 days according to 30-90 [mu]mol, the chicken are normally fed. The prevention and purification feeding method disclosed by the invention has the benefits that the anti-virus ability, the growth rate and the later disease purifying rate of the Wuhua Sanhuang chicken are improved, and the use effect is better.

The invention discloses an avian leukemia resistance molecular marker tva507A > G in a chicken subgroup A and an application of the marker. The invention analyzes the genetic variation of the Chinesechicken species tva gene, and finds that A > G base mutation exists in a 507th position of a Chinese chicken species tva gene DNA sequence (GenBank login number: AY531262.1), and the inventor furtherstudies and proves that the natural mutation of the tva gene can cause the host to generate genetic resistance to ALV-A infection. A detection kit for screening the avian leukemia resistant chickens in the subgroup A is prepared by using the molecular marker, can accurately and quickly determine whether a detection sample is the avian leukosis resistant chickens in the subgroup A or susceptible chickens, and can be applied to screening breeding materials of the avian leukosis genetic resistant chicken varieties (strains) in the subgroup A, and thus breeding of the avian leukemia genetic resistant chicken varieties (strains) in the subgroup A is carried out, so that the marker has good application and popularization value.

The invention discloses a detection method for A, B, J subgroup avian leucosis viruses in an avian infectious laryngotracheitis live vaccine. The detection method comprises the following steps: diluting a commercial avian infectious laryngotracheitis live vaccine by adopting DMEM, then inoculating DF-1 cells, acting for 1 h at the temperature of 37 DEG C, then changing into DMEM containing 1%FBS, and culturing for 7 days; collecting cell culture supernatant, and carrying out avian leucosis virus p27 group specific antigen ALV-p27 detection; collecting cell cultures for an ALV-p27 positive sample and extracting cDNA, identifying A, B, J subgroup avian leucosis viruses by adopting a subgroup specific primer, and carrying out IFA detection on corresponding positive porocytes of the A, B, J subgroup avian leucosis viruses by adopting a subgroup specific monoclonal antibody. The detection method disclosed by the invention can enable live viruses to be proliferated in cells, makes up the defects of direct PCR detection and also has the characteristics of effectiveness, accuracy, reliability and the like and great significance on prevention and control of avian leucosis.

The invention discloses a method for purifying avian leukosis of Rizhao partridge chickens, which relates to the technical field of poultry breeding and comprises the following steps of uniformly arranging specific light sources in a chicken house, and setting specific breeding density, regularly feeding vitamins, selecting a target purified breeding chicken flock, eliminating positive chickens to construct an F0 generation breeding flock, hatching eggs to generate an F1 generation breeding flock, respectively carrying out antigen detection at different times to remove positive chickens, carrying out height, body length and body height detection and screening, constructing an F1 generation breeding purified flock, and hatching eggs to generate an F2 generation breeding flock, and enabling the rest to be done in the same manner until the F4-F6 generation breeding purification population, and acquiring the purification population when the 300-day-old antigen detection reaches zero detection rate. According to the method provided by the invention, population purification of the avian leukosis can be completed, the capability of the population obtained through purification for resisting the infection risk of the foreign avian leukosis is remarkably improved, chickens are not easily infected with the avian leukosis, the morbidity after infection is low, and the death rate after attack is low.