137 results about "Leucosis" patented technology

PCT No. PCT / US96 / 07370 Sec. 371 Date Nov. 28, 1997 Sec. 102(e) Date Nov. 28, 1997 PCT Filed May 22, 1996 PCT Pub. No. WO96 / 37625 PCT Pub. Date Nov. 28, 1996Recombinant avian sarcoma leukosis virus (ASLV)-derived retrovirus vectors having an expanded host range are described. The host range is expanded by the replacement of the ASLV envelope gene by an envelope gene from a virus capable of infecting both mammalian and avian cells. The resulting recombinant ASLV-derived retroviral vectors can replicate efficiently in avian cells, infect both avian and mammalian cells in high titer, and are replication-defective in mammalian cells. Thus, they are quite safe and advantageous for use in gene therapy and vaccines.

The invention relates to the production and use of retroviral vectors for cell specific gene transfer, specially to a production method of retroviral vectors containing capsid particles of murine leukemia virus (MLV) and envelope proteins of human immunodeficiency vises (HIV) or simian immunodeficiency viruses (SIV). Said vectors can be used for gene transfer in selected cell types, specially in CD4-positive mammal cells.

The invention provides a GeXP quick detection kit and detection method for simultaneously identifying 8 chicken infectious immunosuppression disease pathogens. The kit is used on the basis of a GeXP system, and comprises 8 pairs of PCR (polymerase chain reaction) primers. The detection result indicates that the kit has the advantages of high specificity and high sensitivity and can be used for simultaneously identifying and detecting chicken Marek's disease virus, A, B and J subgroup avian leucovirus, avian reticuloendothelium hyperplasia virus, avian reovirus, chicken infectious anemia virus and infectious bursal disease virus.

The invention belongs to the cross technical field of molecular immunology and virology, and particularly relates to a hybridoma cell based on hybridoma type leukosis virus subgroup J isolated strain with preservation number of CGMCC (China General Microbiological Culture Collection) No. 5018, a high-specificity monoclonal antibody of avian leukosis virus subgroup J (ALV-J) surface protein (SU) secreted by the hybridoma cell and a preparation method of the monoclonal antibody. By using the high-specificity monoclonal antibody prepared by the invention, a technical support is provided for diagnosis, prevention and treatment of avian leukosis virus subgroup J disease, and the economic loss caused by avian leukosis virus subgroup J disease is effectively reduced.

The invention discloses an anti-avian leukosis virus p27 protein monoclonal antibody, a gold-colloidal strip containing the same, and application. The anti-avian leukosis virus p27 protein monoclonal antibody is generated by secretion of a hybridoma cell strain with a preservation number of CGMCC NO. 11089 or a hybridoma cell strain with a preservation number of CGMCC NO. 11090. The monoclonal antibody is high in titer to p27 protein and good in specificity, and therefore can be used for preparing a kit and the gold-colloidal strip for detecting an avian leukosis virus. A preparation method of the monoclonal antibody provided by the invention is simple, and an antibody purification process is simple, and the efficiency is high, and the cost is low. As the gold-colloidal strip prepared by the anti-avian leukosis virus p27 protein monoclonal antibody provided by the invention is adopted to detect the anti-avian leukosis virus, the specificity is strong, and the operation is simple, and convenience, speediness and simplicity are realized, besides, a special instrument and equipment are not needed, and professional training is not needed, and a result is clear and easy to recognize; the operation is simple and convenient; the popularization is easy, and the antibody is more suitable for real-time detection.

The invention provides an avian leukosis double-antibody sandwich enzyme-linked immuno sorbent assay (ELISA) antigen detection kit. The kit comprises ELISA plates which are coated on anti-avian leukosis virus (ALV) p27 monoclonal antibodies, enzyme labeling anti-ALVp27 monoclonal antibodies and the like. Antibody capturing and antibody detecting are respectively conducted aiming at different antigenic determinants of p27 protein; anti-ALVp27 monoclonal antibodies coated by the ELISA plates are obtained through secretion of hybridoma cell strain ALVP27-5D3, and the enzyme labeling anti-ALVp27 monoclonal antibodies are obtained through secretion of hybridoma cell strain ALVP27-4F12. The avian leukosis double-antibody sandwich ELISA antigen detection kit is easy, convenient and fast to operate, can be used in detecting of all subgroup virus of ALV, and is suitable for all levels of veterinarian departments in the basic level and rapid and mass screening detecting of the avian leukosis of leaving and entering the country. The avian leukosis double-antibody sandwich ELISA antigen detection kit has the advantages of being low in cost, notable in economical benefit, and wide in application prospect.

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 discloses multiplex PCR (polymerase chain reaction) detection primers for avian leukosis viruses. The primers comprise detection primers for detecting subgroup A, subgroup B and subgroup J of avian leukosis viruses. The inventional also discloses a reaction system and reaction conditions of multiplex PCR. The detection method provided by the invention has the advantages of being rapid, accurate and cheap.

The invention discloses a multiple-connection probe amplification detection kit, primer and probe for simultaneously detecting the bluetongue viruses, the infectious bovine rhinotracheitis viruses, the bovine viral diarrhea viruses, the enzootic bovine leucosis viruses and the foot and mouth disease viruses. The multiple-connection probe is shown in sequence tables from SEQ ID NO:1 to SEQ ID NO:10. The primer is shown in sequence tables from SEQ ID NO:11 to SEQ ID NO:12. The primer, the probe and / or the multiple-connection probe amplification detection kit including the primer and the probe can detect the five vital cow disease pathogenies including the bluetongue viruses, the infectious bovine rhinotracheitis viruses, the bovine viral diarrhea viruses, the enzootic bovine leucosis viruses and the foot and mouth disease viruses at the same time, the detection time and cost are saved, and epidemic diseases can be diagnosed in time.

The preparation of tumor immune cell detection chip and detection method relates to a chip for detecting peripheral blood karyocyte of tumor patient and its detection method. The preparation of its chip includes the following steps: on the carrier making chemical modification, namely modifying upper amino-group and aldehyde group; then on the modified microslide on which the chemical gene is set dropping the correspondent antibody, for example, new protein antibody, etc expressed by gene mutation of tumors of carcinoma of lung, live cancer and carcinoma of stomach and skeletal system tumor leucosis, etc. then placing said microslide in humectous environment and storing, then taking out said microslide and washing by using buffer solution. Besides, said invention also provides its detection method by using said chip.

The invention discloses establishment of a PCR-HRM analysis method for rapid differential diagnosis of different serotypes of avian leukemia viruses. According to the invention, universal primers are good in degeneracy, have very good amplification performance to the A, B, C, D and E subtypes of avian leukemia viruses, is conductive to improving the efficiency of PCR (polymerase chain reaction), and reduces the time of identification and typing of viruses; and specific primers provided by the invention are good in specificity, and the J subtype can be amplified in a specific way. HRM analysis is performed on the amplification product obtained by using the primers, so that the typing of avian leukemia viruses can be performed accurately and rapidly in a high-throughput way by being compared with the standard HRM curve of a known subtype, particularly the endogenous type of avian leukemia viruses and the exogenous type of avian leukemia viruses can be rapidly and accurately distinguished.

The invention provides a method for labeling antibodies by colorful fluorescent granules and a test paper strip prepared from the antibodies. Animal viruses including, but not limited to, canine distemper viruses, canine parvoviruses, canine adenoviruses, canine coronavirus, rabies viruses, feline leukemia viruses, Marek's disease viruses, Newcastle disease viruses and the like are used as targets, and the corresponding antibodies labeled by colorful fluorescent micro-spheres are prepared by the aid of chemical covalent processes and can be applied to detecting the animal viruses. The method for labeling the antibodies by the colorful fluorescent granules and the test paper strip prepared from the antibodies have the advantages that colorful detection lines with bright developed colors can appear during detection, and accordingly qualitative results can be obtained; the contents of the animal viruses in samples further can be subsequently quantitatively obtained, accordingly, test results are clear and are high in stability, and the test paper strip can be stored at the room temperature for a long time.

The invention discloses a typing method of resistance for resisting a subgroup A avian leukosis virus by quality chickens, which adopts an SNP locus at the 619th site of a TVA gene sequence as a research object; PCR-RLFP analysis is performed to detect whether mutation occurs at the SNP locus, and also to detect whether 4 bases are inserted into SNP loci at the 305th-308th sites of the TVA gene sequence; the two detection results are combined together to determine whether the detected chicken is a genetic resistance type or a genetic susceptibility type. The resistance typing method of the invention performs detection from the source, and omits troubles of later-stage detection; the detection method is rapid, accurate, strong in purposiveness, and simple in operation, has practice significance, and can realize rapid selection of chicken genetic characters and improvement of disease resistance of strains.

The present invention discloses an antitumor multi-drug drug-resistant RNA interference drug, provides a group of nucleotide sequences of interference RNA of therapeutic drug pointed at drug-resistant gene mdr-1 and P-glycoprotein (P-gp) expression and function for resisting leucosis and other tumor. It has the target antitumor cell multidrug drug-resistant (mdr-1) gene and P-glycoprotein (P-gp) expression and function. It raises the sensitivity of leucosis cell conventional chemical therapeutic drug and enhances the action of chemical therapeutic drug for killing malignant tumor cell of hemopoietic system.

The invention discloses recombinant J subgroup avian leucosis virus infective cloned plasmids and a preparation method and application thereof. The preparation method comprises the following steps of: replacing long terminal repetitive sequences of two ends of a provirus gene of a J subgroup avian leucosis virus with long terminal repetitive sequences of two ends of a provirus gene of an E subgroup avian leucosis virus; and inserting the long terminal repetitive sequences of the two ends of the provirus gene of the E subgroup avian leucosis virus into vectors, so that the J subgroup avian leucosis virus infective cloned plasmids are constructed, and the obtained sequence is shown as SEQ ID NO:1. The replication capacity and the pathogenicity of recombinant virus strains obtained by transfecting the J subgroup avian leucosis virus infective cloned plasmids are obviously weaker than those of natural J subgroup avian leucosis viruses, but the recombinant virus strains can express structural proteins of the J subgroup avian leucosis viruses, have the antigenicity of the J subgroup avian leucosis viruses and can be identified by a specificity monoclonal antibody JE9 of the J subgroup avian leucosis viruses. The invention provides reference for a material and a method for preparing low-toxicity live vaccines of the J subgroup avian leucosis viruses.

The invention relates to a J subset avian leukosis virus rapid detection test paper card which comprises a lining board; a sample pad, a golden-standard pad, a cellulose nitrate membrane and a water-absorbing pad are disposed on the lining board; the invention is characterized in that the golden-standard pad is coated with colloidal gold labelled monoclonal antibodies of mouse anti-J subset avian leukosis virus gp85 protein; the cellulose nitrate membrane is coated with a detection line formed by rabbit anti-J subset avian leukosis virus polyclonal antibodies and a quality control line formed by rabbit anti-mouse IgG polyclonal antibodies. The advantages of the invention are that: the detection method of the J subset avian leukosis virus rapid detection test paper card of the invention can rapidly detect whether a chicken body is infected by or carries J subset avian leukosis viruses, and has the advantages of rapid reaction, high sensitivity, strong specificity, simple operations, suitability for near-the-pen detection, economy and practicality.

The invention discloses a K subgroup avian leukosis virus detection kit, which relates to the technical field of virus fluorescent quantitative PCR detection. The kit of the invention comprises one pair of specific primers, and nucleotide sequences of the specific primers are respectively as shown by SEQ ID No. 2 and 3. The kit of the invention is simple in application method, low in cost, easy inobserving a reaction result, high in sensitivity, high in specificity, capable of rapidly detecting K subgroup avian leukosis viruses, very suitable for disease monitoring, on-site emergency and clinical sample detection and suitable for large-scale popularization and application.

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 relates to a method for diagnosing avian leukosis virus subgroup J of sicken chicken flocks and belongs to the technical field of molecular biology. According to the method, C type retrovirus characteristics of the avian leukosis virus subgroup J are taken as a breakthrough point, on the theoretical basis of identifying a JL-2 strain of the avian leukosis virus subgroup J, specific primers are utilized to clone gp85 fragments of the virus strain, an expression vector PGEX-6P-1 / gp85 category is constructed, an RFLP method is used to detect the genotype of NRAMP1 genes, the enzyme digestion reaction of the fragments where PCR amplification sites are located is observed, according to the research, it not only is indicated that that cocks carrying T allele have higher resistance to avian leukosis virus subgroup J, but also is found that the specificity diagnosis method established by gp85 is a powerful technical means for detecting specificity of ALV-J and timely purifying populations.

The invention provides a reagent kit for detecting avian leukemia virus J sub-groups (ALV-J), and belongs to the technical field of RT-PCR (reverse transcription-polymerase chain reaction) detection. The reagent kit contains a pair of specific primers. Nucleotide sequences of the specific primers are respectively shown as SEQ ID NO.1-2. The invention further provides a method for detecting the avian leukemia virus J sub-groups. The reagent kit and the method have the advantages of high specificity, sensitivity and efficiency, good universality and low cost. Besides, quick differential diagnosis can be carried out on clinical disease samples in 6.5 h, technical means can be provided for early quick diagnosis on the ALV-J and conduction of molecular epidemiological investigation, and accordingly avian leukemia prevention and control can be effectively guided during poultry raising production.

The present invention provides a water soluble tuckahoe mycelium hetero polysaccharide with anti-tumor activity. By using corn slurry as main component to prepare culture medium, tuckahoe, mycelium is cultured through deep fermentation of wild Tuckahoe strain. Water soluble hetereo polysaccharide is extracted from the mycelium with physiological saline and hot water. The hetero polysaccharide consists of alpha-D-glucose, mannitol, galactose and protein in different content. Experiments shows that the hetero polysaccharides can inhibit the growth of Sarcoma 180 tumor in mouse and acute leukovirus cell, so that they may be used as antitumor medicine and health article for raising body's immunological function.

The invention relates to the field of animal virology and immunology, and provides an epitope vaccine for resisting A / B subgroup avian leucosis virus infection. The epitope vaccine is prepared by highly active recombinant protein His-cENV which is obtained through screening and purification after prokaryotic expression and a freund's adjuvant in a united manner, wherein the nucleotide sequence for encoding the recombinant protein His-cENV is shown as SEQ ID NO.1. Through the epitope vaccine, 7-day-old breeding poultry chicks are immune and can produce 1:128000 neutralizing antibodies. Vitro virus neutralization experiments and animal experiments show that the epitope vaccine can neutralize different ALV-A / B isolated strains, so that chicken flocks are effectively protected to resist the infection of ALV-A / B strains. Because the epitope vaccine is based on a multi-epitope antigen gene sequence which is originated form env, the defect of ALV-A / B virus variation is overcome, a new era of the ALV-A / B vaccine is opened, a new way of resisting the ALV-A / B infection is provided, and a technical support for preventing and controlling the ALV-A / B is provided.

The invention belongs to the technical field of animal epidemic disease serological diagnosis and relates to an avian leukosis P27 monoclonal antibody hybridoma cell strain and application thereof. The avian leukosis P27 monoclonal antibody obtained by secreting the hybridoma cell strain can be applied to detection of the avian leukosis P27, the 96-pore elisa plate is coated by the P27 specific polyclonal antibody, the P27 monoclonal antibody is labeled by alkaline phosphatase, and according to the avian leukosis P27 monoclonal antibody and the P27 monoclonal antibody labeled by alkaline phosphatase, the detection sensitivity, specificity and stability are effectively improved. A high-efficiency and sensitive detection mode is provided for a novel variety which is used for selecting, purifying and breeding avian leukosis positive chickens and has high genetic resistance, and the avian leukosis P27 monoclonal antibody hybridoma cell strain is low in cost, easy and convenient to operate and is suitable for popularization and application of animal husbandry production.

The invention discloses a standard product which is used for detecting the viral load of Friend murine leukemia, and a method which detects the viral load of the Friend murine leukemia by a real-time fluorescent quantitative polymerase chain reaction method (Real-Time RT-PCR method) by using the standard product. The detection process includes the steps of the extraction and content measurement of leukovirus nucleic acid (RNA), the obtaining of the nucleic acid segment by amplifying reverse transcription by using a primer, the detection of the real-time fluorescent quantitative polymerase chain reaction (Real-time PCR), and the like. Compared with conventional PCR detection methods, the method of the invention, which detects the viral load of the Friend murine leukemia, has better specificity. The detected genetic amplified products are target genetic products to be detected. The method has a better linear relationship and is suitable for being applied to the detection of the viral load of the Friend murine leukemia (Fr. MuLV).

The invention discloses a subgroup A or B avian leukosis virus gp85 gene recombinant prokaryotic expression protein and a purification method thereof, and applications. The method includes the following steps: (1) designing and synthetizing specific primers for amplifying gp85 genes; (2) obtaining gp85 gene fragments through PCR amplification, and performing purification; (3) separately cloning the gp85 genes of an ALV-A / B to pET-32a prokaryotic expression vectors, and performing sequencing and identification after screening positive clones; (4) recombining the expression of expression vectorpET-32a-A / B-gp85; (5) purifying expressed gp85 proteins through a urea method; and (6) recombining the renaturation of the gp85 proteins. Thus, soluble gp85 proteins with high purity can be obtained by using the method; and the expressed proteins can be taken as envelope antigens to establish an antibody detection method and used for preparing antiserum for immunizing animals, and also can provideraw materials for immunofluorescence, immunohistochemistry, etc.

The invention relates to the technical field of biological detection, and in particular, relates to a triple fluorescent PCR detection kit and detection method for detection of J subgroup avian leukosis, Marek and reticuloendotheliosis viruses. The detection kit includes a triple fluorescent PCR detection mixed liquid, a negative control and a positive control, wherein the triple fluorescent PCR detection mixed liquid contains three pairs of primers and corresponding probes thereof for amplification and detection of the J subgroup avian leukosis, Marek and reticuloendotheliosis viruses, and the nucleotide sequences are shown in SEQ ID NO:1-9. The kit and the primer probes can quickly detect the J subgroup avian leukosis, Marek and reticuloendotheliosis viruses, and have the advantages of simple operation, high sensitivity, good specificity and the like; and suspected cases can be found out and confirmed in time, and the monitoring level of three kinds of oncosis can be improved.

The invention discloses a gene chip, a kit and a method for detecting three viruses for immunosuppression disease of chickens, and relates to the field of animal medical diagnosis. The gene chip is characterized in that specific probe sequences of four subgroups A, C, D and J of an ALV (Avian leucosis virus) as well as a CAV (Chicken Anemia Virus), and an REV (Reticuloendotheliosis virus) are arranged on a substrate of the chip. The invention further provides the kit for detecting the four subgroups A, C, D and J of the ALV, the CAV and the REV. The chip, kit and detecting method, provided by the invention can be used for detecting various pathogens quickly and accurately in a high-flux manner, so that infectious diseases can be diagnosed correctly within a relatively short period to be prevented from spreading.

The invention relates to a composition containing selfheal and blackberry lily, which comprises the following raw materials, or extracts of the following raw materials or chemically acceptable derivatives of the extracts of the following raw materials in respective dosage: 10 parts of selfheal and 2 to 60 parts of blackberry lily. On the basis, the following one kind or various kinds of but not limited to the following raw materials can be added according to specific conditions: 3 to 50 parts of rheum officinale, 3 to 50 parts of scutellaria baicalensis, 3 to 50 parts of lithospermum, 3 to 50 parts of radices trichosanthis, 3 to 50 parts of radix bupleuri, 3 to 50 parts of osmundaceae cyrtomium fortunei, 3 to 50 parts of honeysuckle and 3 to 50 parts of cassia twig. The composition can be prepared into medicinally or pharmaceutically acceptable physical forms such as powder, liquid, paste, particles, capsules and the like. The composition is used for preventing and treating viral diseases such as bird flu, newcastle diseases, infective bronchitis, infective leukemia, infective bursal diseases, swine fever, parvovirus diseases, epidemic diarrhea, porcine reproductive and respiratory syndrome and the like, and the mortality rate is reduced.

The invention belongs to the technical field of biomedicine and particularly relates to application of seaweed oligosaccharide in the preparation of an avian leucosis virus resistant preparation. seaweed oligosaccharide is prepared by degrading natural polysaccharides extracted from the macroalga Grateloupia filicina, Ulva lactuca, Sargassum fusiforme and other algae, has a wide source range, is simple to prepare, and has the advantages, as an antiviral preparation, such as naturalness, zero toxicity and environmental friendliness; in-vitro cell experiments prove that the seaweed oligosaccharide is capable of inhibiting the proliferation of avian leucosis virus, has significant antiviral effect, may be applied to livestock and poultry breeding as a novel antiviral preparation, and has high applicable value.

The invention discloses an infectious cDNA (complementary deoxyribonucleic acid) clone, a construction method and application of a recombinant subgroup J avian leucosis virus capable of expressing EGFP (enhanced green fluorescent protein). The infectious cDNA clone has a nucleotide sequence shown by SED ID NO.1. The infectious cDNA clone of the recombinant subgroup J avian leucosis virus capable of expressing the EGFP can be used for successfully rescuing viruses. Moreover, the rescued recombinant subgroup J avian leucosis virus capable of expressing the EGFP can not influence the replication of the virus after the insertion of extraneous source EGFP, overcomes the complexity in the conventional process of detecting the an ALV-J neutralizing antibody at the same time, and can be used for large-scale detection of the ALV-J neutralizing antibody more simply and effectively.