230results about How to "Promote amplification" patented technology

The invention discloses a method for simultaneous and efficient amplification of CD<3+>CD<56+>CIK cells and CD<3->CD<56+>NK cells. The method comprises the steps as follows: the concentration of separated PBMC (peripheral blood mononuclear cells) is adjusted by a serum-free medium containing autologous plasma, an Anti-CD16 antibody, IL-2 and IL-15 are added, and then the mixture is transferred into a T175 culture flask for culture; an Anti-CD3 antibody and an Anti-CD137 antibody are added; a serum-free medium containing the autologous plasma, IL-2 and IL-15 is supplemented every two days according to the cell growth condition; the cell concentration is controlled to be about 1.5*10<6> / ml; and after culture is performed for 14-21 days, large quantities of high-purity CD<3+>CD<56+>CIK cells and CD<3->CD<56+>NK cells can be obtained simultaneously, and the total cell quantity can reach an effective value of the cell quantity required for adoptive cellular immunotherapy clinically for tumor. The method for simultaneous and efficient amplification of the CD<3+>CD<56+>CIK cells and the CD<3->CD<56+>NK cells is simple, convenient, effective and high in cell killing activity.

The invention provides a preparation method of freeze-dried fruit, vegetable, nut and cereal yoghourt nutritive cubes. The preparation method comprises the following steps of cleaning fruits and vegetables, performing pulping, mixing and blending pulp with yoghourt, performing homogenizing, adding cereals, nuts and fruit granules, performing stirring, performing membrane filling, performing quick-freezing, performing demolding, performing vacuum freeze drying, and performing packaging so as to obtain the freeze-dried fruit, vegetable, nut and cereal yoghourt nutritive cubes. The freeze-dried fruit, vegetable, nut and cereal yoghourt nutritive cubes are produced by a vacuum freeze drying technique, so that original thermal sensitivity components and antioxidation components of products canbe sufficiently guaranteed, the color and the fragrance of fruits and vegetables and the activity of probiotics can also be guaranteed, and transportation and long term storage are facilitated; and besides, the cereals, the nuts and the fruits cooperate, so that the nutrition is more balanced.

The invention discloses an expansion method of various lymphocyte subpopulations and application of the expansion method to preparation of an adoptive immunotherapy medicine for cancers. The expansion method comprises the following steps: S1, adding IFN gamma and zoledronic acid into a culture medium of PBMC collected in the blood of a tumor patient, and re-adding an OKT3 antibody and recombinant human interleukin-2 after culture to stimulate PBMC so as to finish preliminary expansion; S2, after finish of preliminary expansion, mixing immune lymphocytes with feeder layer cells, and after adding zoledronic acid, an OKT3 antibody and recombinant human interleukin-2, continually performing expansion. By adoption of the expansion method, after twice expansion, the sum of the immune cells can be 10,000 to 80,000 times by expansion, the expanded immune cells include alpha beta T cells, gamma delta T cells, NKT cells and NK cells, and tumor cells can be directly killed, or a cellular immunotherapy drug is prepared to kill the tumor cells.

The invention discloses a method and a device for detecting and selecting laser-induce enhanced Raman spectrum in a liquid. The method comprises the following steps of: focusing the inducing light on a suspending to-be-detected sample and forming a three-dimensional light well in liquid; enriching metal nanoparticles on the surface of the to-be-detected sample; and coinciding the focuses of exciting light and inducing light, thereby forming a laser-induce enhanced Raman signal. The realizing method disclosed by the invention is simple and convenient, is obvious in spectrum enhancing effect and is fit for quick detection for in-situ Raman spectrum of a biological sample under a liquid environment; the to-be-detected sample and the metal nanoparticles are hatched together; the method is especially fit for the field of quick detection for diseases, epidemic situation, food safety, and the like; the toxic damage to the biological sample caused by the metal nanoparticles is greatly reduced, so that the true physiological status of living cell can be extremely reflected; the optical tweezers formed according to the method provided by the invention can be used for sorting, extracting and eluting, can realize the function of identifying and sorting Raman spectrum of a single living cell, and is beneficial to the follow-up study on the biological sample.

The invention discloses a method and a kit for simultaneously constructing a sequencing library by DNA and RNA. The method comprises the following steps: (1) carrying out reverse transcription on RNAin a reaction substrate to form RNA / cDNA composite double strands; (2) directly adding Tn5 transposase and a fragmentation buffer solution into a mixture of a reverse transcription product cDNA / RNA composite double strand and a DNA double strand for fragmentation, and adding a part of linker sequence to the 5' end of the double strands during fragmentation; (3) strand displacement and library amplification: adding an amplification buffer solution and a strand displacement and amplification enzyme mixture into the obtained product, and carrying out PCR amplification; and (4) carrying out magnetic bead purification to obtain a sequencing library of the original DNA / RNA. According to the method, the library building steps are greatly reduced, the library building time is shortened, the library output and offline data quality is improved, and more real information is helped to be obtained.

The invention discloses a method and kit for realizing multiple detection of nucleic acid through a colloidal gold chromatographic technology, and belongs to the field of medical biotechnology. The invention provides a nucleic acid detection test strip, a universal probe is marked with colloidal gold particles to be fixed on a glass cellulose film, the universal probe is designed into a universal sequence, an NC film on the test strip have one or more detection lines and a quality control line, each detection line is coated with a section of nucleic acid sequence which can be in specific hybridization combination with a corresponding specific probe B series; the quality control line is coated with a section of nucleic acid sequence which can in specific hybridization combination with a specific probe A series; the specific probe A series is in complementary pairing hybridization with the universal probe; the probe marked with gold and a nucleic acid amplified fragment are combined in series by virtue of the specific probe A series and the specific probe B series, so that the specific detection of the nucleic acid fragment can be realized. The method has the advantages that the technical requirements of experimenters are low, the required detection time is short, special instruments are not required, and the method is easy to popularize towards grass-roots and remote countryside medical institutions.

The invention provides an expansion in vitro purification culture method of mesenchymal stem cells and a cell culture medium used in a culture process. According to the method, mesenchymal stem cells differentiate to osteogenic stem cells. The cell culture medium comprises the following components: 8-12mu g / L of bFGF (Basic Fibroblast Growth Factor), 8-12mu g / L of EGF (Epidermal Growth Factor), 8-12mu g / L of PDGF-BB (Platelet Derived Growth Factor-BB), 10 percent of fetal calf serum, 100U / mL of penicillin and 100mg of L of streptomycin; a solvent is a basic culture medium; and the basic culture medium is a DMEM (Dulbecco Modified Eagle Medium) culture solution, an F12 culture solution or a mixture of the DMEM culture solution and the F12 culture solution. The mesenchymal stem cells obtained by adopting a method comprising the steps of early replacement of the cell culture medium, cell adherence passage and combined addition of cell factors (bFGF, EGF and PDGF-BB) in the culture solution are high in purity and strong in vitro proliferation capacity, and have good capability of differentiation to osteogenic stem cells.

The invention relates to the technical field of gene sequencing, and discloses a biological chip detecting system. The system comprises a PCR (polymerase chain reaction) reaction device, a hybridization device, a reading device and a transportation device, wherein the PCR reaction device is used for augmenting samples in a PCR pipe; the hybridization device comprises a reaction chamber and a chipbearing table for holding a biological chip to enable the samples to hybridize with the biological chip; the reading device is used for detecting the biological chip treated by the hybridization device and obtaining relevant gene information; the transportation device is used for transporting the PCR pipe to the PCR reaction device and transporting augmented samples to the hybridization device from the PCR reaction device or transporting a hybridized biological chip between the hybridization device and the reading device. The transportation device is used for automatically transporting the PCRpipe, the samples in the PCR pipe and the hybridized biological chip, the integration and systematization of the PCR reaction device, the hybridization device and the reading device are realized, themechanization and automation of the biological chip detection are realized accordingly, the manual operation is reduced, so that errors brought about by the manual operation are reduced, and the detection efficiency of the biological chip is improved.

The invention discloses a sequencing library construction method and kit for pathogenic microorganism detection. The method comprises: (1) putting an extracted sample into a sample preservation solution, carrying out DNA / RNA co-extraction, and removing host rRNA; (2) adding reverse transcriptase to enable RNA in the sample to be subjected to reverse transcription to form RNA / cDNA composite doublestrands; (3) directly adding mutant Tn5 transposase and a breaking buffer solution into a mixture of the reverse transcription product cDNA / RNA composite double strand and a DNA double strand for fragmentation, and after the fragmentation is finished, carrying out a termination reaction by using a termination reaction solution 5*TS-plus; (3) adding an amplification buffer solution and a strand displacement and amplification enzyme mixture, and carrying out PCR amplification; and (4) carrying out magnetic bead purification to obtain a sequencing library of the original DNA / RNA. According to themethod, library building steps are greatly reduced, the library building time is shortened, the requirement on the initial quantity of samples is reduced, and the library output and offline data quality are improved.

The invention provides an erythrocyte lysis buffer. The erythrocyte lysis buffer comprises the following components: 4.5-5.5 mmol / L of sodium chloride, 0.8-1.3% (v / v) of triton 100, 300-340mmol / L of glucose and 0.008-0.012 mol / L of trishydroxymethyl aminomethane. The erythrocyte lysis buffer can effectively lyse the erythrocyte in a whole blood sample, separate and enrich leucocyte in the blood, and facilitate the subsequent nucleic acid extraction operation of the leucocyte, so as to obtain the leucocyte DNA in the whole blood sample and the DNA of virus in the leucocyte.

The present invention provides one kind of specific EST-STS marking primer-1 of rye 1RS chromosome. The marking primer-1 is designed with the EST sequence, BE637935, of the first partial wheat homologous group as the template, and has the sequences including STSWE126F of 5'-TCAAGCACGCATTTCAACTC-3' and STSWE126R of 5'-ACAGATGTCCAAAGCCCAAC-3'. The primer STSWE126 has one amplified 850 bp size specific segment located in rye 1RS chromosome. The marking primer-1 of the present invention expands the use range of wheat EST, provides one new tool for deep research of distant hybrid material for common wheat. The marking primer-1 may be applied in identifying rye 1RS chromosome simply and fast by means of common PCR technology.

The invention discloses a lymphocyte culture fluid and culture method thereof and application therefor, the culture fluid is adding ATP and coenzyme A in completely culture fluid containing basic culture fluid RPMI-1640, albumin, L-glutamine, glucose, baking soda and HEPES. The culture method includes steps: (1) preparing culture fluid; (2) suspending cell in culture fluid prepared by step (1); (3) activating cell; (4) washing activated cell by the culture fluid prepared by step (1); (5) collecting cell. The culture fluid of the invention cultures cell with higher amplification times and longer generation time.

The invention discloses a KIR and ligand genetic typing experimental method. By the technology, the time and labor are saved, and meanwhile, DNA sample capacity is further saved. A multi-PCR technology is adopted, meanwhile, 36 pairs of primers are further combined according to the sizes of PCR products, and finally, amplified reaction is finished in 12 reaction holes. The KIR and ligand genetic typing experimental method is conveniently applied to amplification of 96 pore plates, conventional Taq enzyme is used, typing of all KIR genes and ligands thereof can be finished at a time under the same reaction conditions, and the practicality of the KIR and ligand genetic typing experimental method is greatly improved. Two pairs of primers are used for a KIR gene, the accuracy is improved, anda false negative result is avoided. The KIR gene can be combined to MHC-I type ligand molecules on the surfaces of targeting cells, inhibiting or activating signals are transmitted to regulate the activity of NK cells and T cells, and the KIR and ligand genetic typing experimental method plays an important regulation role in hematopoietic stem cell transplantation, feto-matemal tolerance, anti-infectious immunity, tumor immunity and autoimmune diseases. Therefore, KIR genetic typing facilitates understanding of influences of KIR to tumor immunity, hematopoietic stem cell transplantation and autoimmune diseases.

The invention relates to the field of cells, and discloses a culture medium for mesenchymal stem cells and a large-scale culture method thereof. The culture medium for mesenchymal stem cells disclosed by the invention comprises a serum-free culture medium, EGF and PDGF, wherein the concentration of the EGF is 5-10ng / mL, and the concentration of the PDGF is 5-10ng / mL. The large-scale culture method for mesenchymal stem cells disclosed by the invention comprises the following steps: taking P1-generation mesenchymal stem cells which are primarily isolated and cultured, adjusting an inoculum density to be 2*10<4> / mL to 3*10<5> / mL by the culture medium for mesenchymal stem cells, culturing to a cell fusion degree of 80-90% and then recording as P2-generation, and repeatedly passing to P10-generation cells. The culture medium disclosed by the invention reduces a cell pollution probability, promotes MSC amplification, and delays the ageing speed of MSC in in-vitro culture. The large-scale culture method disclosed by the invention is capable of obtaining lots of high-purity mesenchymal stem cells, and keeping the good stem cell characteristics of the mesenchymal stem cells.

The invention discloses a method for co-culturing cord blood stem cells and mesenchymal stem cells, belonging to the biotechnology and tissue engineering technical field. The method is characterized by adding no blood serum but only cytokine and trophocyte; wrapping the trophocyte calcium alginate-chitosan gel beads; using microcarrier and rotating wall vessel to implement the co-culture of cord blood stem cells and mesenchymal stem cells; and separating and obtaining the cord blood-derived stem cells respectively. The invention has the advantages that: the microcarrier provides a surface for the cord blood mesenchymal stem cells to adhere to, so the adherent, dynamic, suspension culture of the cord blood mesenchymal stem cells can be implemented. The rotating wall vessel provides an environment for hemopoietic stem cell to suspend and growth in, and also reduces the damages caused by fluid shear stress to the hemopoietic stem cells. The throphocyte can nourish the cord blood stem cells outside the gel beads, replace partially blood serum and contribute to the separation and harvest of different cells.

The invention discloses a 3D printed Ti-PDA-PLGA microsphere bone defect repair stent. A 3D printed Ti stent is prepared by a laser sintering technology. Then, under certain conditions, dopamine is self-polymerized on the fiber surface of the 3D printed Ti stent to form a PDA coating, thereby preparing a 3D printed Ti-PDA stent; then PLGA microspheres carrying VEGF are prepared by a double emulsion-solvent evaporation method, and finally, BMP-2 and PLGA microspheres carrying VEGF are adsorbed and immobilized on the surface of the stent by an adsorption method, and finally the 3D printed Ti-PDA-PLGA microsphere bone defect repair stent is formed. The bone defect repair tissue engineering stent disclosed by the invention has the advantages of reliable mechanical property, high biological activity and safety, convenient implantation, small trauma and low cost, and can be used for the repair treatment of bone defect after bone traumas, bone tumors and bone infection.

The invention discloses a separation and authentication method for denitrifying bacteria with anaerobic ammoxidation activity, wherein a novel anaerobic ammonoxidation bacterium segregation and evaluation program is employed, wherein denitrifying bacteria is isolated first, then the capability of ammonia and nitrite simultaneous transformation under anaerobic condition is used as the criterion for confirmation of anaerobic ammonoxidation bacterium, finally the classification status of the bacterium is identified.

The invention relates to a serum-free cryopreservation medium and method for peripheral blood monouclear cells. The serum-free cryopreservation medium comprises recombinant human interleukin-2, humanserum albumin, polyethylene glycol, trehalose and a basic culture medium or sodium chloride for injection. The serum-free cryopreservation medium has the advantages that the recombinant human interleukin-2 (IL-2) is added into the cryopreservation medium, so that the activity stability of immune cells cultured by thawed cells can be increased greatly, the high activity of original cells can be kept, the immune cells can well keep the physiological function and biological feature of the thawed cells, and the problem of direct large-scale amplification of the thawed cells can be solved effectively; the polyethylene glycol and the trehalose are added into the cryopreservation medium, the cryopreservation medium can replace an existing dimethyl sulfoxide-based (DMSO-based) cryopreservation medium, the toxicity of the cryopreservation medium to the cells is lowered effectively, cell stability is maintained, the direct application safety of the thawed cells is increased, and the thawed cellscan be directly used.

The invention discloses a drug metabolic enzyme related gene SNP fluorescence labeling composite amplification kit. The kit comprises a mixture of specificity non-labeled primers and share labeled primers of five SNP loci, performs detection by combining PCR composite amplification with a capillary electrophoresis technology and can simultaneously detect genotypes of the five SNP loci of a drug metabolic enzyme related gene in a single tube within 3 hours. By applying the kit, the drug metabolic enzyme related gene is detected, the relation of the adverse drug reaction and genetic variation is researched, and a theoretical support is provided for rational drug use.

The invention discloses a 3D printed PCL-PDA-BMP2 bone tissue engineering scaffold and a preparation method thereof. The 3D printed PCL-PDA-BMP2 bone tissue engineering scaffold is prepared accordingto the following steps: adopting a fusion extruding forming type 3D printing technology for extruding PCL and forming fiber bundles and preparing a 3D printed PCL scaffold through a splicing frameworkof the fiber bundles; forming a PDA coating by automatically polymerizing dopamine on the fiber surface of the 3D printed PCL scaffold, thereby preparing a 3D printed PCL-PDA scaffold; lastly, soaking the acquired 3D printed PCL-PDA scaffold in a BMP2 solution, thereby acquiring the 3D printed PCL-PDA-BMP2 bone tissue engineering scaffold. The scaffold prepared according to the invention has theadvantages of simple and reliable structure, controllable appearance and microstructure, reliable mechanical properties and controllable ion release property, and can be used for restorative treatmentof bone trauma, bone tumors and bone defect after bone infection.

The invention discloses a Taq DNA polymerase mutant and application thereof. The Taq DNA polymerase mutant has amino acid substitutions at one or more of the following amino acid positions in the sequence shown as SEQ ID NO. 1; and the various amino acid substitutions are represented in triplets as 'letters-numbers-letters', wherein the numbers reflect position of an amino acid mutation, the letters before the numbers are corresponding to an amino acid involved in the mutation, and the letters after the numbers reflect an amino acids used to replace the amino acid corresponding to the lettersbefore the numbers. The Taq DNA polymerase mutant disclosed by the invention is capable of changing configuration of an enzyme, thereby improving tolerance of the enzyme; and thus, the mutant can be very well applied in blood sample amplification.

The invention discloses a 3D printed Ti-PDA-PLGA microsphere bone defect repair stent and a preparation method thereof. A 3D printed Ti stent is prepared through a laser sintering technology; then, under a certain condition, dopamine is self-polymerized on the fiber surface of the 3D printed Ti stent to form a PDA coating, so that the 3D printed Ti-PDA stent is prepared; and then VEGF-carrying PLGA microspheres is prepared by a multiple emulsion solvent evaporation method, and finally the BMP2 and the VEGF-carrying PLGA microspheres are absorbed and fixed on the surface of the stent by an adsorption method, finally the 3D printed Ti-PDA-PLGA microsphere bone defect repair stent is prepared. The bone defect repair tissue engineering stent of the invention has the advantages of reliable mechanical property, high biological activity and safety, convenience in implantation, small trauma and low cost, which can be used for repairing treatment of bone trauma, bone tumor and bone defect afterbone infection.

The invention discloses a 3D printed PCL-Mg bone tissue engineering scaffold and a preparation method thereof. The 3D printed PCL-Mg bone tissue engineering scaffold is prepared from poly-epsilon-caprolactone PCL and magnesium chloride as raw materials through a 3D printing technology. After being implanted into the human body, PCL as a bioabsorbable material is gradually degraded and releases Mgto promote osteochondral generation. The porous structure of the scaffold can induce bone ingrowth and finally repair bone defects, tumors and bone defects after infection. The 3D printed PCL-Mg bonetissue engineering scaffold has the advantages of simple and reliable structure, controllable shape and microstructure, reliable mechanical property, controllable ion release performance, convenient implantation, small trauma and low cost.

The invention relates to techniques for detecting drug resistance mutation of Mycobacterium tuberculosis and provides a method and kit for detecting ethambutol resistance mutation of Mycobacterium tuberculosis, which have the advantages of simple design, simplified operation, high analysis sensitivity and low cost. The method comprises the following steps of: extracting DNA of a Mycobacterium tuberculosis sample; designing primers and probes by using the primer design software PrimerPremier 5 according to the embB gene sequences of Mycobacterium tuberculosis, constructing a PCR (polymerase chain reaction) system, performing PCR, performing melting curve analysis and detection result judgment, and determining whether the mutation occurs in the regions which are covered by the probes according to the Tm change of the analyzed melting curve. By adopting three pairs of primers and four probes and constructing the double-tube double-color system, the purpose of detecting the mutations of six codons 306, 368. 378, 380, 406 and 497 on the embB gene is achieved.

The invention discloses an umbilical cord blood Treg cell in-vitro amplification method based on trophoblastic cells and application. The specific technical method comprises the steps that firstly, umbilical cord Wharton's jelly mesenchymal stem cells are adopted as the trophoblastic cells to induce preliminary proliferation of Treg cells in umbilical cord blood mononuclear cells; then, pure Tregcells are obtained through magnetic bead sorting; and finally, the Treg cells are stimulated to be rapidly amplified by using optimized amplification factors. According to the amplification method, human AB plasma, IL-2, rapamycin, an RARA agonist and a DNA methyltransferase inhibitor are used as the optimized amplification factors, and a large number of umbilical cord blood Treg cells with high purity and high activity can be prepared within two weeks. Umbilical cord blood is used as a raw material for Treg cell amplification, batch preparation can be achieved, and Treg cell quality fluctuation caused by individual differences of samples can be reduced. The umbilical cord blood Treg cells have low immunogenicity and can be used as universal cells for clinical research, such as autoimmunediseases, graft-versus-host diseases and the like.

The invention relates to a biofloc culture method and an aquatic culture method. The biofloc culture method comprises the following steps of adding an organic silicon source into a water body, wherein the adding amount of the organic carbon source meets the requirement that a C / N ratio in the water body is (15 to 17):1; adding bacillus licheniformis, bacillus laterosporus and lactobacillus casei into the water body, wherein the total adding amount of the bacillus licheniformis, the bacillus laterosporus and the lactobacillus casei is 0.8*10<5> to 1.2*10<5>cfu per 1ml of water body; aerating the water body, and maintaining dissolved oxygen in the water body to be 6mg / L or more. The biofloc culture method has the advantage that by adding probiotics into the water body, and adjusting the adding amount of the organic carbon source in the water body, the certain C / N ratio in the water body is maintained, mass propagation of the probiotics in the water body is effectively promoted, the formation of biofloc is accelerated, and the large-size biofloc is obtained.

The invention relates to the technical field of cell culture, in particular to a building method of NK (natural killer) cell and gamma delta T cell co-culture capable of realizing the in vitro commonmultiplication culture on NK cells and gamma delta T cells. The method mainly comprises the following steps of S1, reagent selection; S2, performing PBMC culture by a culture medium for 3 days; S3, supplementing the culture medium; maintaining the concentration; performing culture for 4 days; S4, after the seventh day, removing supernatant through centrifugation; transferring the cells into a T25cell culture bottle; then, replacing an EX culture medium; S5, after the tenth day, supplementing the culture medium for maintaining the concentration; S6, after the twelfth day, continuously supplementing the culture medium for maintaining the concentration; S7, after the fourteenth day, detecting the NK cell and gamma delta T cell proportion and the cell total number. The co-culture building method has the advantages that two kinds of anti-tumor immune cells with similar properties are cultured in one step; the cell culture efficiency is improved; the culture cost can be greatly reduced through being compared with that of independent culture of various immune cells for obtaining NK cells and gamma delta T cells; the culture technical difficulty is also reduced.

The invention belongs to the field of crop breeding, and relates to a molecular marker of wheat scab resistance expansion gene Fhbl and application thereof. The molecular marker is one of MAG6067 and MAG6660 with a genetic distance to an Fhbl gene being respectively 0.05cM and 0cM. A molecular marker which is closest linked to the Fhbl for the first time internationally by utilizing the invention. The Fhbl gene can be detected by marking the MAG6660 to determine whether the Fhbl exists or not and the existing state of the Fhbl, and the scab resistance of wheat can be predicted, so that a plant carrying the Fhbl can be quickly screened; and the molecular marker can be used for breeding a scab-resistance variety. Furthermore, laboratory detection can be carried out by utilizing the molecular marker to avoid the influence of the environment to the phenotype. According to the molecular marker closely linked with the Fhbl, time and labor for screening scab resistance materials can be greatly saved, the prediction accuracy can be enhanced, and clone of the Fhbl gene can be accelerated.

The invention discloses a nested fluorescence reverse transcription-polymerase chain reaction (RT-PCR) detection method for avian influenza virus (AIV) H5 subtype with high efficiency and high sensibility and a detection kit. In the method, two sets of polymerase chain reaction (PCR) primers (a pair of outer primers and a pair of inner primers) are utilized to carry out PCR amplification twice, that is, a product obtained after the outer primers are subjected to PCR amplification is taken as a template which is used for carrying out the PCR amplification on the inner primers; a binding site of the inner primers and the template DNA is located at the inner side of a DNA fragment which is obtained by amplifying the outer primers; the nested PCR is very effective in reducing or eliminating nonspecific amplification and improving sensitivity. Compared with common detection methods, the method disclosed by the invention has good specificity, and can exactly detect the subtype AIV of H5N1 and H5N2 and detect H1N1, H3N2, H6, H9NDVLasata and EDSV to be negative; is very beneficial to amplifying an AVI micro-template in a fish farming water body, and can completely meet the requirements onsensibility and specificity of AVI detection in the cultivation water of fishponds.

The invention relates to exosomes derived from granulocytic myeloid-derived suppressor cell and an application thereof, belongs to the fields of cytobiology, molecular biology and clinical application and particularly discloses a tumour individual or autoimmune disease individual (exosomes) (called G-MDSC exo) derived from the granulocytic myeloid-derived suppressor cell and application in preparation of medicine for inhibiting the autoimmune disease. The invention finds that the G-MDSC exo can effectively inhibit the CD4+T cell proliferation effect in vitro, promote the T regulartory cell induced proliferation and relieve the tardive hypersensitivity model mice foot swelling degree; the G-MDSC exo has inhibiting effects on the mice inflammatory bowel disease (IBD) and the collagen-induced arthritis (CIA). The application in preparation of medicine for effectively treating the autoimmune disease is also disclosed.