102 results about "Chick embryos" patented technology

The invention relates to a preparation method and a product of an H9N2 subtype avian influenza inactivated vaccine. The technical points of the invention mainly relate to the screening, the determination and the domestication of a virus-adapted cell line, the primary amplification cultivation and the continuous cultivation of a virus-adapted cell, the preparation of virus fluid by virus-inoculated culture and the preparation of final inactivated vaccine products. Firstly, the invention avoids the virus propagating method using a large amount of chick embryos in the avian influenza production at present, thereby avoiding the problem of biological potential safety hazards, and overcoming the problem that the mass production of vaccines is enslaved to the supply of the chick embryos; secondly, the invention provides a safe, continuous and closed cell culture virus production method, is used for the preparation of the H9N2 subtype avian influenza inactivated vaccine, enables the use of the cell culture method, and can simultaneously produce high-titer viruses to meet the requirements for the immunological production; and finally, the vaccine production method of the invention is simple and fast, thereby realizing the fast vaccine supply at the epidemic situation.

The invention discloses a gender identification method for a chick embryo in a near-infrared hatching egg at earlier stage of incubation, and belongs to a hatching egg detection technology. The method comprises the following steps: obtaining a near-infrared spectrum of a hatching egg through an optical fiber probe and a Fourier near-infrared spectrometer in a dark room, decomposing the near-infrared spectrum of the hatching egg by an overall mean empirical mode, removing high-frequency characteristic mode components to remove noise, removing low-frequency characteristic mode components by a method based on empirical mode decomposition to achieve baseline correction, extracting main components of spectral data as input variables of a nerve network so as to finish the identification, wherein the output value of the nerve network is 0 or 1. According to the method, the gender identification efficiency and accuracy of the hatching egg at earlier stage of incubation are high, the incubation cost is reduced, and the development of layer chicken and meat chicken feeding production is facilitated.

The invention discloses a genotype VII Newcastle disease virus marker vaccine strain and an application thereof, and belongs to the field of genotype VII Newcastle disease virus marker vaccine strain rescue and application. A built Newcastle disease virus reverse genetic operating platform is utilized for enabling NP protein of a G7 strain to miss 18 amino acids and conducting mutation on F-protein cleavage loci, and the highly-weak virulence and high-virus titer genotype VII Newcastle disease virus marker vaccine strain MG7-NPdelta18+Fmut is rescued through screening. The microbial preservation serial number is CCTCC NO: V201505. The marker vaccine strain has the biological characteristics of high growth titer and low virulence in chick embryos and is genetically stable. The immune protection test result shows that the marker vaccine strain is good in immunogenicity, capable of inducing high-level protective antibodies, and capable of completely protecting immunized chicken, can be used for preventing and controlling a currently-popular genotype VII Newcastle disease virus and lays the foundation of identifying vaccine immunity and wild virus infection.

The invention aims at providing a new castle disease and H9 subtype bird flu bivalent vaccine. The new castle disease and H9 subtype bird flu bivalent vaccine contains antigens and adjuvant. The antigens are inactivated H9 subtype bird flu viruses and new castle disease viruses. The H9 subtype bird flu viruses are QDY strains, and the preservation number of the H9 subtype bird flu viruses is CCTCC v201517. The QDY strains of the H9 subtype bird flu viruses and a Lasota strain of the new castle disease viruses are inoculated to chick embryos respectively, and then virus liquid is collected; the virus liquid and the oil adjuvant are mixed and emulsified into the vaccine after the virus liquid is inactivated through a formaldehyde solution. The new castle disease and bird flu bivalent inactivated vaccine is good in immunogenicity, antibody production is fast after immunity, the produced antibody titer is high, the produced antibody holding time is long, the retention period is long, the immunizing dose is small, the selected adjuvant is easy to inject, and two kinds of diseases can be prevented through one-time injection. The vaccine has the advantages of being efficient and good in safety.

An improved process for recovery of virus from allantoic fluid of virus-infected chick embryos. Virus associated with granular and fibrous debris in the allantoic fluid can be disassociated from the debris and recovered, thereby increasing viral yield. Dissociation can be achieved by subjecting the virus-debris complex to conditions of increased salt concentrations, e.g., 0.5 M or greater.

The invention provides a preparation method of a muscovy duck original goose parvovirus and duck type II adenovirus combined inactivate vaccine. The preparation method is characterized in that targeted process conditions are designed according to the proliferation characteristics and immunogenicity of the muscovy duck original goose parvovirus and the duck type II adenovirus. Concretely, chick embryos are used for proliferating the muscovy duck original goose parvovirus; LMH cells are used for proliferating adenovirus; after being inactivated, the viruses are prepared into safe and effective muscovy duck original goose parvovirus and the duck type II adenovirus combined inactivate vaccine. When being used, the prepared muscovy duck original goose parvovirus and duck type II adenovirus combined inactivate vaccine has the advantages that the antibody generation is early; the protection rate is high. The vaccine can be used for preventing two diseases after once inoculation; the stress response caused by multi-time vaccine inoculation of the muscovy duck can be solved.

The invention relates to a Newcastle disease chimeric virus marker vaccine strain as well as a construction method and application thereof, and belongs to the field of rescue and application of Newcastle disease chimeric vaccine strains. A Newcastle disease virus reverse genetic operation platform is utilized to mutate an F protein cleavage site of a Newcastle disease gene GVII type strain into acleavage site of an attenuated strain, F and HN genes of a mutated gene VII type Newcastle disease strain and NP, P, M and L of a gene II type NDV La Sota strain construct a full-length chimeric cDNAsequence, and a 18bp nucleotide marker sequence is inserted into a non-coding region between P and M. A Newcastle disease chimeric virus NDV DC strain is obtained through transfection cell rescue. Theconstructed chimeric virus can reach relatively high culture titer in chick embryos and cells. The chimeric strain contains envelope surface glycoprotein of the gene VII type Newcastle disease strain, contains a skeleton of the gene II type strain, and has immunogenicity of the Newcastle disease gene VII type strain and high reproduction and high safety characteristics of the gene II type La Sotastrain.

The invention relates to an acquisition method and adaptive sites of an influenza A virus vaccine mammalian cell adaptive strain. According to the invention, a conventional influenza A vaccine skeleton strain is subjected to continuous passage in mammalian cells so as to obtain the mammalian cell adaptive strain; four amino acid sites on the viral protein coded by a virogene in the adaptive strain undergo mutation, which contributes to adaptability to the skeleton strain. The adaptive strain does not has enhanced pathogenicity on living bodies like chick embryos and mice, but has substantially improved adaptability to mammalian cells.

The invention relates to applications of a BHK-21 cell full-suspension culture technology in production of a newcastle disease vaccine. A process of producing the newcastle disease vaccine by BHK-21 cell full-suspension culturing includes steps as follows: (1) a step of viral strain seed selection, namely a step of inoculating a monolayer BHK-21 cell with a newcastle disease vaccine virus seed cultured by a chick embryo, adding a virus maintenance medium, culturing to obtain a newcastle disease vaccine adapted to the BHK-21 cell, and performing system identification; (2) a step of domestication and seed selection of a suspension cell strain, namely a step of domesticating a full-suspension BHK21 cell used for culturing of the newcastle disease vaccine virus and establishing a basic seed; (3) a step of subjecting the suspension cell to enlarged cultivation; (4) a step of virus inoculation and harvest, namely a step of inoculating the newcastle disease vaccine virus adapted to the BHK-21 cell and harvesting a virus solution; and (5) a step of measuring the viral titer of the multiplicated newcastle disease vaccine virus and preparing the vaccine. According to the applications, culturing and production with chick embryos of the newcastle disease vaccine are changed into to large-scale culture and production with mammalian cells of the newcastle disease vaccine, the process of producing the newcastle disease vaccine is simplified, the production cost is reduced, and the yield and quality of the vaccine are largely improved.

The invention belongs to the technical field of medicines and in particular relates to a preparation method of chick embryo extract. Chick embryos are percolated and extracted by utilizing alcohol liquid; a production process is relatively simple; the disadvantage that effective components are damaged by high-temperature extraction is overcome and the disadvantages that a previous process is complicated, low-temperature operation is not easy to control and requirements on equipment are high are also overcome. The chick embryo extract can be used for a traditional Chinese medicine prescription or is singly used; the extract provided by the invention is used for resisting fatigue and improving the immunity and has the advantages that the bioavailability is high and the extract is suitable for large-scale mass production.

The invention relates to a cell extraction method in the technical field of bioengineering, in particular to an extraction method for chick embryo myoblasts. The method includes the following steps that firstly, chick embryos are sterilized, and pectoralis major is taken and put into a D-Hank's culture dish; secondly, the pectoralis major is trimmed; thirdly, the pectoralis major is oscillated and suspended through a vortex oscillator; fourthly, filtration and centrifugation are carried out; fifthly, cell plate laying and differential attachment are carried out; sixthly, digestion is carried out to obtain the myoblasts; seventhly, counting and plate planting are carried out. By means of the extraction method for the chick embryo myoblasts, namely, the mechanical oscillation extraction method for the chick embryo myoblasts, it is guaranteed that the purity, number and vitality of the extracted myoblasts are higher than those of myoblasts extracted through other methods, and a foundation is laid for further development of poultry muscle development regulation and control mechanism experiments.

The invention belongs to recombinant virus carrier vaccines in the field of molecular biology and biotechnology, and particularly relates to a recombinant turkey herpesvirus virus strain rHOH expressing H7N9 subtype avian influenza virus haemagglutinin protein and a construction method. The preservation number of the recombinant turkey herpesvirus virus strain rHOH is CGMCC NO.12985. A gB promoter of the recombinant turkey herpesvirus virus strain rHOH comes from a turkey herpesvirus virus carrier FC126 itself; an expressed heterologous antigen (haemagglutinin protein) gene is optimized through codon bias of Gallus Gallus, the nucleotide sequence changes, and no amino acid sequence changes. The recombinant turkey herpesvirus virus strain rHOH can be used for preparing vaccines for preventing H7N9 subtype avian influenza viruses; 18-day-old chick embryos obtained after hatch or 1-day-old chicks are inoculated, an H7N9 epidemic situation of poultry is prevented and controlled, and public health and safety are protected.

The invention discloses a method for preparing a nutritious intestinal mucosa repair agent for animals by separating and extracting chick embryo EGF, as well as an application of the nutritious intestinal mucosa repair agent for the animals. According to the method for preparing the nutritious intestinal mucosa repair agent for the animals, epidermal growth factors (EGF) are separated and extracted from rapidly growing and developing chick embryos; and the method specifically comprises the following steps of adjusting pH value of a homogenate solution so as to make EGF with positive charges, adsorbing the EGF by using water-insoluble benzoic acid so as to separate the EGF from the homogenate solution, and resolving the EGF from the benzoic acid by utilizing acetone which has a property of EGF insolubility so as to prepare a chicken-source-EGF-containing nutritious repair agent which is capable of promoting intestinal mucosa repair of the animals by adopting the principle design. The nutritious intestinal mucosa repair agent for the animals disclosed by the invention is added into full-value feeds for livestock and poultry at an addition amount of 0.05-0.3 kilograms / ton; and the added nutritious intestinal mucosa repair agent is capable of obviously improving structural functions of the intestinal mucosa of the animals, increasing feed intake and growth performance of the livestock and poultry, and significantly reducing diarrhea rate of weaned piglets.

The invention provides a method for detecting a special growth and development law of muscles of Jingning chicken. The Jingning chicken are unique and peculiar local chicken of the Gansu province, are fresh and tender in texture and unique in flavor, and have good public praise, however, the Jingning chicken are endangered, and the prospect is grim. In the method for detecting the special growth and development law of the muscles of the Jingning chicken, chick embryos of the Jingning chicken are researched, and special meat quality trait controlling genes such as growth and development gap-associated proteins: CECR2 (cat eye syndrome chromosome region) and IGF-1 (insulinlike growth factor-1), fatty acid metabolism gap-associated proteins: Ex-FABP (fatty acid-binding protein) and LPL (lipoprotein lipase) and protein degradation gap-associated proteins: CAST and CAPN1 are selected. Protein expression distribution and change of expression level of proteins in posterior limb buds and body segments in a chick embryo development HH19-31 period are detected by using methods of immunohistochemistry and Western blotting and the like, and the Jingning chicken are compared to white feather broilers, yellow feather broilers and hyline brown chicken. By the method for detecting the special growth and development law of the muscles of the Jingning chicken, the relation of growth and development regulatory functional genes, growth performance and meat quality is discussed, and data are provided for establishing target genes of transgenic chicken. Moreover, the resource superiority can be widely developed and used, and new vitality is injected to development of local economy.

The invention provides a method for separating acian metapneumovirus, which adopts tracheas and lung tissues mixed cells of specific pathogen free (SPF) chick embryos which are incubated for 17-19 ages in days to cultivate the acian metapneumovirus. The method for separating the acian metapneumovirus successfully improves acian metapneumovirus (AMPV) vitro culture conditions through a separation and reproduction method of a same culture system and two kinds of mixed cells based on a traditional method that a clinical development sample is separated and cultured into a kind of sensitive cell at one time, obviously improves separation rate of AMPV, and reduces reproduction times.

The invention discloses a recombinant equine influenza virus strain, a preparation method thereof and a vaccine prepared from the recombinant equine influenza virus strain. The recombinant influenza virus strain contains genes HA and NA of an equine influenza virus A / equine / xinjiang / 3 / 07 (H3N8) strain and six internal genes PB2, PB1, PA, NP, M and NS of an influenza virus A / Puerto Rico / 8 / 34 / Mount Sinai (H1N1) or A / PR / 8 / 34 (H1N1 short for PR8 virus). The recombinant equine influenza virus strain disclosed by the invention is named as rH3N8-PR and is preserved with the number of CGMCC NO.8161. The invention also discloses a preparation method of the recombinant equine influenza virus strain and a vaccine prepared from the recombinant equine influenza virus strain. Compared with a parental strain, the recombinant equine influenza virus strain disclosed by the invention can generate very high virus titer and blood clotting titer on both chick embryos and MDCK (Madin Darby Canine Kidney) cells, and the pathogenicity of the recombinant equine influenza virus strain to mice is remarkably reduced; experiments prove that the vaccine prepared from the recombinant equine influenza virus strain disclosed by the invention has favorable immunogenicity and protective effect.

The invention discloses an antiaging nutrient composition. The antiaging nutrient composition is prepared from nucleotide, lysine, methionine, phenylalanine, threonine, tryptophan, arginine, histidine, glycine, aspartic acid, leucine, isoleucine, valine, serine, glutamine, glutamic acid, proline, tyrosine, cysteine, gamma-aminobutyric acid, taurine, orotic acid, granulesten, egg yolk lecithin, cephalin, alpha-linolenic acid, gamma-linolenic acid, inositol, hydroxytyrosol, vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin E, niacin, folic acid, iron, zinc, manganese, copper, selenium, chromium, potassium, calcium, magnesium, choline, L-carnitine, pentose, sodium carbonate and freeze-dried powder of chick embryos hatched for three days. The antiaging nutrient composition has the advantages that neural stem cell proliferation is accelerated, amount of autophagosome is increased and activity of mesenchymal stem cells telomerase is improved.

The invention relates to a virus isolate for infectious bronchitis and application in vaccine preparation. The virus isolate is named as an isolate cb2, and has been collected into Common Microorganism Center of China Committee for Culture Collection of Microorganisms on June 21st, 2016; the microorganism collection number is CGMCC No.12657. The virus isolate has the advantages that the isolated strong-poisonous virus isolate for the infectious bronchitis is used for the adaptive domestication on chick embryos, so as to obtain the virus isolate with high titer for the growth of the chick embryos; a stable disease occurrence model can be established, the good specificity and immunogenicity of the virus isolate are maintained, and the virus isolate can be used for the production and preparation of vaccines; after the virus isolate is deactivated, the virus isolate is prepared into a deactivating vaccine to perform evaluation and testing on the immune effect and safety; proofed by results, the vaccine is safe to the targeted animal, the adverse effect of local and whole body due to vaccines is avoided, and the infection caused by kidney and respiration type infectious bronchitis viruses can be effectively prevented.

The invention discloses a preparation method of antioxidant activity mixed peptide originated from chick embryos, belonging to the technical field of bioactive peptide extraction. The preparation method comprises the following steps of: collecting healthy chick embryos which are incubated for certain days, and extracting a mixed peptide finished product with the antioxidant activity from the chick embryos through machining methods of homogenating, freezing, drying, re-dissolving with sterile water, centrifuging at low temperature, filtering by using an ultra-filtration membrane, freezing, drying and the like, so as to obtain a mixed peptide finished product with antioxidant activity is extracted from the chick embryos. The finished product can be added into food or cosmetics as functional factors, or is developed as an ingredient of a novel, natural and efficient antioxidant medicine. The method is simple, the cost is low, the obtained product has the advantages of nature, health, high activity and the like, and particularly the product has potential development values for the middle aged or the young in sub-health; and meanwhile as the natural anti-oxidative peptide is applied to health-care food and cosmetics, the consumption requirements of people on origin and pure nature are met. The preparation method is also a high additional value machining method of the chick embryos, and is an application of the food-medicine homology theory in traditional Chinese medicine in the health-care field.

The invention provides an H5 and H7 subtype avian influenza virus genetic engineering subunit vaccine, and a preparation method and application thereof. The extracellular region of H5 and H7 subtype avian influenza virus HA protein is captured to construct a shuttle vector containing an HA extracellular region gene, an sf9 insect cell is transfected, a recombinant virus is saved, through an hi5 cell, secretory expression is carried out, purification is carried out through affinity chromatography to obtain an HA protein antigen, and an adjuvant is added and emulsified to prepare the H5 and H7 subtype avian influenza virus genetic engineering subunit vaccine. The preparation method disclosed by the invention is simple, overcomes the defects generated by producing vaccines by chick embryos, is short in time consumption, is high in an expression quantity, can realize cell suspension culture in a reactor, and is favorable for large-scale production. The prepared genetic engineering subunit vaccine has a good immune effect, can effectively prevent infection of H5 and H7 subtype avian influenza viruses and has a wide application prospect.

The invention provides a pigeon paramyxo virus type 1 PPMV-1 / BJ-C strain and application thereof. The attenuated strain PPMV-1 / BJ-C is constructed by mutating an F protein cleavage site of the attenuated strain PPMV-1 / BJ-C into a La Sota strain corresponding site on the basis of a pigeon plague virus virulent strain PPMV-1 / BJ. PPMV-1 / BJ-C has the advantages of good immunogenicity, good culture characteristics, stable titer and the like, the pigeon plague virus PPMV-1 / BJ-C strain is inoculated to 9-11-day-old SPF chick embryos through an allantoic cavity, allantoic fluid of infected chick embryos is harvested and inactivated with diethyleneimine, and then an aluminum hydroxide adjuvant is added to prepare a pigeon plague virus inactivated vaccine. The inactivated vaccine prepared from the PPMV-1 / BJ-C strain can generate a relatively high antibody, can prevent ND caused by pigeon plague virus, and the vaccine has the advantages of safety, quick response, lasting immune period and the like, and has wide application prospects.

The invention discloses a method for preparing a flu vaccine. The method comprises the following steps of: a) performing adaptability passage on the original viruses of flu viruses through unspecific pathogenic chick embryos to serve as main-generation seeds; b) preparing primary hamster kidney cells, and culturing by using a culture flask or a cell biological reactor; c) infecting the by using the main-generation seeds, and performing adaptability passage until viruses with the virus clotting titer of not lower than 1:640 are obtained and used as working seeds of the vaccine; d) infecting the primary hamster kidney cells by using the working seeds, performing virus amplification until vaccine monovalent stock solution with the virus clotting titer of not lower than 1:320 is obtained; ande) concentrating, inactivating and purifying the vaccine monovalent stock solution, mixing different types of monovalent virus solution, and sub-packing into finished products. The primary hamster kidney cells have adequate sources and are easy to produce on a large scale. The prepared vaccine does not contain reproducible deoxyribonucleic acid (DNA) which has tumorigenicity on human bodies or animal bodies and has high safety.

The invention discloses a chicken infectious bronchitis divalent live vaccine and a preparation method thereof. The divalent live vaccine comprises antigens and a freeze-drying protective agent. The antigens comprise a chicken kidney-type infectious bronchitis virus K136 strain having an accession number of CCTCC-V201320 and a chicken breath-type infectious bronchitis virus H120 strain. The preparation method includes respectively performing amplification culture to the IBV-K136 strain and the H120 strain through chick embryos; mixing culturing products uniformly to prepare the vaccine antigens; and mixing uniformly the vaccine antigens and the freeze-drying protective agent and performing freeze drying to prepare the vaccine. The divalent live vaccine can prevent the chicken breath-type infectious bronchitis and the chicken kidney-type infectious bronchitis simultaneously. The vaccine conforms to the epidemiological characteristics of the chicken infectious bronchitis in our country at present, and solves a problem that no vaccine for the chicken kidney-type epidemic infectious bronchitis is available in our country at present. The divalent live vaccine can be popularized and used in a large scale.

The invention relates to a chimeric Newcastle disease virus vector H7 (avian influenza) live vaccine candidate strain capable of overcoming an effect of a Newcastle disease virus maternal antibody of a chick and a construction method thereof. The preservation number of the Newcastle disease vaccine candidate strain is CGMCC No.13798. The construction method includes: making use of a reverse genetic manipulation platform of a chimeric Newcastle disease virus rAI4-TFHN which is capable of being replicated in the existence of a Newcastle disease antibody, inserting an HA (hemagglutinin) sequence of a subtype H7N9 (avian influenza) virus strain into a full-length transcription vector of an AI4-T4FHN strain genome so as to obtain a full-length cDNA (complementary deoxyribonucleic acid) clone pNDV / rAI4-T4FHN-H7 of a recombinant Newcastle disease virus genome containing the subtype HA gene of the H7N9 (avian influenza) virus. The recombinant virus AI4-T4FHN-H7 obtained through transfection is high in propagation titer in chick embryos, still capable of expressing HA protein stably after continuous passage and applicable to large-scale production of vaccine and can be used for manufacturing the vaccine.

The invention discloses a method for rapidly preparing a large number of chick embryo fibroblasts. The method comprises steps as follows: 1) taking of chick embryos; 2) pancreatin washing; 3) homogenization; 4) digestion; 5) dispersion; 6) subculture, and obtaining of the chick embryo fibroblasts. The yield of the chick embryo fibroblasts obtained with the method is more than 20 times that of chick embryo fibroblasts obtained with traditional methods, and the operation time is short. Practices prove that each chick embryo can be transferred in 10-20 T225 cell bottles.

The invention discloses a high-yield influenza virus preparation system and a preparation method, on one hand, MDCK cells lacking MARCH8 are used for preparing influenza viruses, the virus productiontiter is improved, and an influenza virus preparation system based on mammalian cells is optimized; on the other hand, high-yield strain with M2 mutation, which is not affected by MARCH8, is used forpreparing influenza viruses, so that the yield of the influenza virus prepared in mammalian cells and chick embryos is increased, and the method for preparing the influenza virus by utilizing the mammalian cells and the chick embryos is optimized.

The invention discloses a nano influenza vaccine, a construction method and application. The nano influenza vaccine is a recombinant protein 3M2e-rHF and has a sequence shown in SEQ ID No. 2. According to the recombinant protein disclosed by the invention, 3M2e is displayed on the surface of a cage structure of ferritin, so that the immunogenicity of M2e is remarkably improved, and thus, a novel influenza vaccine, i.e., nanoparticles 3M2e-rHF is constructed. According to the nano influenza vaccine, the construction method and the application, a recombinant protein vaccine is expressed by using a procaryotic expression system, i.e., Escherichia coli, and no live virus is involved in a vaccine preparation process, so that compared with the traditional methods for preparing influenza vaccines from chick embryos, the method has the advantages of being safer, simpler and more convenient in operation and being suitable for carrying out rapid large-scale production; by using the high sequence conservation of the M2e in different subtype influenza viruses, proven by experiments, the 3M2e-rHF protects mice from resisting the infection of homotype and heterotype influenza viruses, thereby being advantageously developed into general influenza vaccines with cross protection potency.

By improving the seed virus diluent, adding aspartic acid, ZnCl2 and epidermal growth factors in a specific proportion on the basis of a phosphate buffer, so that the H7 subtype avian influenza virusin the low-immunity chick embryo can be efficiently proliferated. The level of virus proliferation and the immune effect of the prepared vaccine are basically close to those of SPF chick embryos and non-immune chick embryos, however, the production cost is effectively reduced, the production threshold is lowered, which has a positive significance in production of H7 subtype avian influenza vaccines.