34 results about "Hematopoietic organ" patented technology

The invention discloses a method for preparing a rainbow trout IHN (Infectious Haematopoietic Necrosis) inactivated vaccine, which comprises the following steps: carrying out grinding, filtering and poison dipping processing on pancreas and livers of juvenile fish which is attacked, but is still alive, inoculating rainbow trout gonad (RTG) cells, carrying out blind passaging at 14 DEG C, keeping for 5 days when carrying out blind passaging on the tenth generation, and collecting cell poisonous fluid; inoculating chinook salmon embryonic (CHSE) cells, carrying out passaging at 14 DEG C, and raising the culture temperature by 1 DEG C when passaging for 5 generations each time until the culture temperature is raised to 20 DEG C; and adopting epithelioma papulosum cyprini (EPC) cells to continuously carry out passaging under the condition of 20 DEG C, passaging to the twelfth generation to obtain high-titer virus solution and carrying out inactivation to obtain the rainbow trout IHN inactivated vaccine. According to the preparation method, RTG-2, CHSE-214 and EPC cells are utilized to alternately culture rainbow trout IHN viruses at a specific environment temperature so as to obtain a high-titer IHNV (Infectious Hematopoietic Necrosis Virus) antigen and produce the inactivated vaccine; and the technical difficult problem that the high-titer IHNV antigen cannot be obtained through single cells is solved.

An inducing agent or enhancing agent, for the expression of HM1.24 antigen in hematopoietic tumor cells, comprising interferon α, interferon γ, or the IRF-2 protein as an active ingredient, as well as an anti-tumor agent for hematopoietic tumors which comprises a combination of said inducing agent or enhancing agent and an antibody against HM1.24.

The invention discloses an infectious hematopoietic organ necrosis vaccine and a method for amplifying a virus of the infectious hematopoietic organ necrosis vaccine on muscle cells of Pimephales promelas. The invention provides the method for proliferating infectious hematopoietic organ necrosis virus in vitro. The method comprises the following steps of: inoculating the infectious hematopoietic organ necrosis virus to muscle cells of Pimephales promelas according to the MOI value of 0.001, culturing, and collecting supernate, so as to proliferate the infectious hematopoietic organ necrosis virus on the muscle cells of Pimephales promelas. According to the present invention, the IHNV is inoculated to the FHM cell at the concentration of MOI = 0.001, the required virus collection time is short, and the virus titer is high and stable. According to the proliferation scheme, viruses are amplified on a large scale, the inactivated vaccine is prepared from BPL and formaldehyde, the result shows that the immune effect of the vaccine on rainbow trout is good, and the relative immune protection efficiency can reach 80% or above.

The invention discloses RT-LAMP detection primer pairs, a detection kit and a detection method for infectious haematopoietic necrosis virus (IHNV). The detection primer pairs comprises a pair of outer primers, a pair of inner primers and a pair of loop primers; the detection kit comprises primer liquid, reaction liquid, DNA polymerase, reverse transcriptase and control; and the detection kit also can contain a color developing agent. The detection method comprises the following steps: extracting to-be-detected virus RNA, and amplifying a sample RNA template at the temperature of 63-65 DEG C by utilizing reverse transcription activity of reverse transcriptase and adopting six specific primers and one DNA polymerase with strain displacement activity, wherein the pg grade of pure virus RNA can be detected; and identifying by adding SYBR Green I ESE-Quant-tube Scanner instrument detection, or utilizing a turbidity meter for observing change of turbidity of sediments in a reaction tube, so as to judge whether to carry out amplification or not and determine whether the to-be-detected sample contains IHNV RNA or not. The RT-LAMP detection primer pairs, the detection kit and the detection method for the IHNV have the advantages of quickness, high efficiency, easy operation, high specificity, high sensitivity, easy identification and applicability to field detection and are applicable to popularization and application.

The invention discloses a preparation method for a standard sample of an infectious hematopoietic organ necrosis virus molecule. The preparation method is characterized by including following steps: extracting an infectious hematopoietic organ necrosis virus RNA (ribonucleic acid); performing RT-PCR (reverse transcription-polymerase chain reaction); purifying target genes; performing connection transformation of target segments; recycling plasmid; performing in vitro transcription; performing RNA purification after transcription; and performing phenol chloroform extraction. The preparation method is time-saving and labor-saving in a preparation process, the standard sample of prepared infectious hematopoietic organ necrosis virus ribonucleic acid is high in stability and fine in uniformity, standard samples can be provided for infectious hematopoietic organ necrosis virus detection researches, medicine researches, application researches and the like, so that comparison of different laboratory results is realized, and quality control of laboratories is guaranteed.

The invention discloses an infectious hematopoietic necrosis vaccine and a method for amplifying infectious hematopoietic necrosis virus (IHNV) on epithelioma papulosum cyprini (EPC). The invention provides a method for preparing an infectious hematopoietic necrosis vaccine, and the method comprises the following steps: 1) inoculating IHNV to EPC according to an MOI value of 0.0001, carrying out culturing, and collecting supernatant, thereby obtaining proliferated IHNV; and 2) preparing the infectious hematopoietic necrosis vaccine by using the proliferated IHNV. Experiments in the invention prove that when the IHNV is inoculated to the EPC with the concentration of MOI = 0.0001, the required virus harvest time is short, and the virus titer is high and stable. The IHNV is amplified on a large scale based on the proliferation scheme, and an inactivated vaccine is prepared from BPL and formaldehyde. Results show that the vaccine has a good immune effect on Oncorhynchus mykiss, and the relative immune protection efficiency was up to 84%.

The invention provides an inactivated vaccine for an epizootic haematopoietic necrosis disease in carassius auratus and a preparation method of the inactivated vaccine for the epizootic haematopoieticnecrosis disease in the carassius auratus. The vaccine comprises spinal cord tissue cell lines of carassius auratus gibelio and cyprinidherpesvirus II, and a preservation number of the spinal cord tissue cell lines of the carassius auratus gibelio is CCTCC NO: C2018211. The preparation method comprises the following steps: performing CyHV-2 virus amplification on the cultured spinal cord tissue cell lines of the carassius auratus gibelio to obtain CyHV-2 virus solutions; and performing inactivation treatment on the CyHV-2 virus solutions. The vaccine prepared by adopting the method disclosedby the invention has a good immunoprotection effect, can be applied to preventive immunity for the epizootic haematopoietic necrosis disease in the carassius auratus, and can increase survival ratiosof raised carassius auratus and the raising efficiency of the carassius auratus.

The invention discloses a compound medicine and a method for preventing and treating infectious haematopoietic necrosis of masu salmon, and belongs to the technical field of aquaculture. The medicineis prepared from the following raw materials in parts by weight: 17%-23% of honeysuckle, 15%-25% of radix scrophulariae, 12%-18% of wild chrysanthemum flower, 14%-16% of melia azedarach leaf, 7%-13% of bighead atractylodes rhizome, 5%-15% of dried ginger, 3%-7% of houttuynia cordata and 2%-8% of liquorice. The specific prevention and treatment method comprises the following steps: aeration of culture water, immersion bath of iodine methionine and disinfection of the culture water, and vitamin C, vitamin E and compound medicines need to be added into the daily ration feed at the same time. Themethod is simple to operate, does not influence the quality of aquatic products, and is widely applied to prevention and treatment of infectious hemopoietic necrosis of masu salmon.

The invention discloses a high-efficient PCR (polymerase chain reaction) detection method of infectious hematopoietic necrosis virus and relates to a detection method of the infectious hematopoietic necrosis virus. The invention aims at the problem that the existing PCR method for inspecting the infectious hematopoietic necrosis virus is not high in sensitivity. The method comprises the following steps: 1) designing specific primers IHNV-Lf and IHNV-Lr; 2) getting a tissue filtrate after treatment of a detected pathological material; 3) preparing RNA (ribonucleic acid) of the IHNV (infectious hematopoietic necrosis virus); 4) performing PCR amplification to get an amplified product; and 5) observing the PCR amplified product and determining a result to end the detection. The detection method disclosed by the invention has great specificity, and the cross reaction with VHSV (viral hemorrhagic septicemia virus) does not exist; and according to the detection method disclosed by the invention, a polymerase protein is taken as a detection target, the specific primers IHNV-Lf and IHNV-Lr are utilized for detection, the operation is simple, the accuracy is high, the detection method is sensitive, the sensitivity under the condition of low concentration is great, the detection cost is relatively lower, and the detection method is more time-saving in comparison with an existing nested PCR (two-round PCR).

The invention discloses yang-strengthening and nourishing liquor and a preparation method thereof. The liquor includes alcohol (ethanol) and also includes extracts of bone, blood and kidney of animals and extracts of traditional Chinese herbal medicines, wherein the liquor includes the main raw materials of fresh collected bone, blood, fresh kidney, traditional Chinese herbal medicines capable of nourishing yin, strengthening yang, dispelling wind and activating blood, and grains or bran. The liquor satisfies the theory of "tonifying viscera by means of viscera" in traditional Chinese medicines. Kidney stores essential substances, governs fluid and bones and engenders marrow; bone contains large amount of minerals and organic substances and is one of hematopoietic organs; blood contains various proteins, glucose and other nutritional substances and belongs to iron-containing substances. By means of the extracts of the bone, blood and kidney, the liquor has the effects of soothing the meridian to free the collaterals, improving body vitality and strengthening yang and nourishing yin. The raw materials of the liquor are wide in sources and are low in cost, so that the liquor is low in production cost and has simple production process, and can be produced not only industrially but also domestically.

The invention relates to a method for detecting fish contagious hemopoietic organ necrosis virus based on RPA and belongs to the technical field of biodetection. The method comprises the following steps: determining RPA primers according to a virus G protein conserved sequence, an upstream primer: 5'-GATGAGTGGGAGGGCTTTCACGGATTGC-3' and a downstream primer: 5'-GCACGCGGCAACAGCAAGGAGGAGAACAAGG-3'; extracting virus RNA from an RNA extraction kit box, preparing cDNA from virus RNA through an inverse transcription kit and freezing the cDNA for later use; detecting impurity and content of cDNA undera nucleic acid protein analyzer according to OD260/OD280 and concentration value; preparing an RPA reaction system; and identifying an RPA product. The method has the beneficial effects that the specific primers are designed according to a gene G of an IHNV conserved sequence, and the detection method is simpler and more efficient, high in specificity and high in sensitivity, is not dependent on ahigh end instrument, is suitable for field rapid detection, and can control burst of fish epidemic diseases.

The invention relates to the field of nucleic acid detection, especially disclosing a primer for detecting necrosis virus of goldfish hematopoietic organ by loop-mediated isothermal amplification, a kit based on the primer and a method for detecting necrosis virus of goldfish hematopoietic organ by loop-mediated isothermal amplification. The primer disclosed by the invention is selected from at least one group of 1-5 primer groups designed according to polymerase gene DNA of the necrosis virus of goldfish hematopoietic organ, helicase gene, terminase gene and triplex protein gene. The method and primer for detecting necrosis virus of goldfish hematopoietic organ by loop-mediated isothermal amplification disclosed by the invention provide a new detection method of good sensitivity and strong specificity for necrosis virus of goldfish hematopoietic organ, which can be completed just at constant temperature, is simply operated and specially suitable for inspection and quarantine departments as well as field inspection.

The invention discloses a method for increasing blood storage protein (SP) amount of silkworm larvae in an early stage. Positions corresponding to hematopoietic organs of the silkworm larvae in a target stage are exposed to through holes of a separation plate, and the silkworm larvae are stimulated by heavy ion beams or laser irradiation through the through holes. When the heavy ion beams stimulation is carried out, the dose is controlled at 20-40 Gy. When the laser irradiation stimulation is carried out, the irradiation time is 20-80 s. The method better solves the problem that the amount ofthe blood SP of the silkworm larvae is less in the early fifth instar. The left and right sides of second and third thoracic segments of the silkworm larvae (where the hematopoietic organs exist) arephysically stimulated to induce the mass production of the SP in the early fifth instar of the silkworm. The induction treatment can increase the blood SP-1 content by 15-21% and the blood SP-2 content by 14-17%.

The invention discloses an RAA constant-temperature fluorescence detection method and detection kit for epidemic hematopoietic organ necrosis virus (EHNV). The detection kit comprises a forward primerSEQ ID NO. 1, a reverse primer SEQ ID NO. 2, a specific fluorescent probe SEQ ID NO. 3, a reaction solution, reverse transcriptase, recombinant polymerase and a reference substance. The kit of the present invention has high specificity; the detection sensitivity is high, and can reach 6 fg/[mu]L; the kit has high accuracy and reliability; and the kit has simple and fast operation, is suitable forfield detection, and has wide application scenarios.

A medicament for improving prognostic survival in therapy of malignant tumor is provided that may improve prognostic survival in DIC patients where the basal disease is malignant tumor, especially malignant tumor in hematopoietic organs. The medicament according to the invention comprises as a main active ingredient Activated Protein C, which is obtained from plasma or prepared by using the genetic recombination technique, and efficiently prolongs life-span of DIC patients where the basal disease is malignant tumor, especially malignant tumor in hematopoietic organs. In particular, the medicament may reduce adverse side effects of chemotherapeutics in chemotherapy of malignant tumor to enhance efficacy of said therapy and improve prognostic survival of patients suffering from malignant tumor.

The present invention discloses a set of reagents for detection of infectious pancreatic necrosis virus. The kit for detecting infectious pancreatic necrosis virus disclosed in the invention consistsof six single-stranded DNAs named B2-F3, B2-B3, B2-FIP, B2-BIP, B2-LooF and B2-looB; B2-F3, B2-B3, B2-FIP, B2-BIP, B2-LooF and B2-looB are respectively SEQ ID No. in the sequence listing. Single-stranded DNA as shown in 1-6. The experiment proves that the kit of the present invention has good specificity for detecting infectious pancreatic necrosis virus, and does not cross-react with infectious hematopoietic organ necrosis virus (IHNV) and viral hemorrhagic septicavirus (VHSV); sensitivity is high, sensitivity is 0. .0008fg / 25[mu]L reaction system; detection is simple and time-saving.

The invention belongs to the field of extracts, and particularly relates to preparation and application of an anti-IHNV kelp extract. The invention provides a method for preparing a kelp extract, which comprises the following steps: 1) preparing a kelp crude extract: removing protein in a kelp leaching solution to obtain the kelp crude extract; the kelp leach liquor is an extracting solution obtained by carrying out enzymolysis and water extraction on kelp, and 2) the kelp extract is obtained by carrying out ethanol precipitation on a dialysis retention solution and collecting precipitates, and the dialysis retention solution is in-bag liquid obtained by carrying out dialysis purification on the kelp crude extract by using a dialysis bag method. The invention further provides application of the kelp extract in preparation of an infectious hematopoietic necrosis virus inhibitor or an infectious hematopoietic necrosis virus resisting medicine. The inhibition rate of the kelp extract with the concentration of 300 mu g/mL to IHNV reaches 63.44%.