74 results about "Echinococcosis" patented technology

The present invention relates to the field of antigen gene technology, specifically to an echinococcosis antigen gene, namely egG1Y162 antigen gene and its recombinant protein and applications, wherein, the gene has a nucleotide sequence with a sequence 1. The invention obtains the echinococcosis antigen gene, namely egG1Y162 antigen gene and its recombinant protein from the granule echinococci, performs cloning, transforming and induce expression to the egg1y162 gene, the animal experiment further indicates that the egg1y162 recombinant protein vaccine has a protective effect on the infection of secondary granule echinococci to mice, becomes a candidate gene vaccine for preventing and controlling echinococcosis, and provides a new way for the development of practical vaccine.

The invention discloses a sheep echinococcosis ELISA (enzyme-linked immunosorbent assay) antibody detection kit. The detection kit comprises a sheep echinococcosis recombinant antigen Eg95 pre-coated ELISA plate, a sample diluent, a positive reference solution, a negative reference solution, an enzyme bonder, a scrubbing solution, a substrate color-developing solution and a stop solution. A preparation method of a sheep echinococcosis recombinant antigen Eg95 comprises the steps as follows: firstly, PCR (polymerase chain reaction) amplification is performed on an Eg95 gene segment, then EcoRI and XhoI are used for performing enzyme digestion on a PCR product and a prokaryotic expression vector pET-32a simultaneously, a target segment and the vector after gel recovery are connected to construct a recombinant plasmid pET-EG95, and the recombinant plasmid is transformed into Escherichia coli, induced by IPTG (isopropyl-beta-d-thiogalactoside) for expression and purified by Ni-NTA. According to the kit, recombinant protein Eg95 is used as the antigen to detect a sheep echinococcosis antibody, and the antibody detection kit is high in specificity, high in sensitivity and good in stability.

The invention discloses an imaging model in small animal living bodies with echinococcus granulosus and a construction method thereof. The construction method comprises the steps of: feeding CF-1 male mice; carrying out in vitro culture and drug treatment on protoscolex; carrying out in vitro fluorescence imaging and fluorescence gradient analysis; pretreating mice with depilatory paste for experimental animals, injecting the abdominal cavity of each mouse with chloral hydrate anesthetic, implanting three groups of stained scoleces under Glisson capsules by injection, dividing the mice into three groups according to different groups of injected protoscolex; in a control group, injecting the livers of mice with protoscolex on which drug treatment is not carried out, collecting images every 12 hours after injection, carrying out fluorescence intensity analysis on the ROIs region, putting into a polyethylene cage after imaging, and carrying out in vitro fluorescence gradient detection in the positive control group. The construction method constructs a mouse model with echinococcus granulosus which can express luciferase, provides a platform for drug sensitive tests in living bodies with echinococcosis, and can monitor the growth and transfer of hydatid in the living model dynamically in a long term.

The invention discloses an echinococcosis serum Raman spectrum diagnostic apparatus based on an optimal back-propagation neural network. A laser Raman spectrometer scans each sample twice and is usedfor acquiring serum samples of healthy people and echinococcosis patients, acquiring spectrum data corresponding to the samples, averaging the spectrum data to serve as Raman spectrum data of the samples and transmitting the data to a computer. The computer receives the Raman spectrum data, Raman fluorescent backgrounds in the Raman spectrum data are eliminated by an adaptive iteration penalized least-square method, and noise of the Raman spectrum data without the fluorescent backgrounds is removed by a self-normalization algorithm. The computer takes 3 front main components as new variable input space for constructing a sample matrix after dimension reduction of the Raman spectrum data without the fluorescent backgrounds and after noise removal by a partial least-square method, classification is performed by the back-propagation neural network, 74 samples are selected among 123 samples and used for a training set, and 49 samples are used for a testing set.

The invention discloses a preparation method of sheep echinococcosis infection-resistant gene engineering subunit vaccine. The preparation method comprises the following steps: S1, searching and downloading EG95 gene sequence as shown in SEQ ID NO.2 as well as EG95 amino acid sequence as shown in SEQ ID NO.3 of echinococcus granulosus from NCBI and performing amino acid sequence modification to obtain the modified EG95 amino acid sequence; S2, performing tandem expression on the gene sequences of a plurality of modified EG95 amino sequences through flexible linker to form recombinant multi-EG95 gene sequence; S3, cloning the recombinant multi-EG95 gene sequence to a pET28b plasmid vector, converting to Escherichia coli BL21 (DE3), and performing induction expression by a tag added fusion expression mode to obtain recombinant protein; and S4, performing protein purification on the recombinant protein to obtain the sheep echinococcosis infection-resistant gene engineering subunit vaccine. According to the preparation method, the production cost of echinotype antigen can be greatly reduced, the production process is greatly simplified and various advantages of safety, high efficiency,low cost and the like are achieved.

The invention discloses a vaccine for blocking transmission of echinococcosis pathogeny echinococcus granulosus from a source. The vaccine provided by the invention is a CTB-EgM123 vaccine or a complete set of vaccine consisting of the CTB-EgM123 vaccine and a subunit vaccine of an EgM123 protein recombinant mycobacterium smegmatis vaccine. An active ingredient of the CTB-EgM123 vaccine is a fusion protein formed by fusion of a cholera toxin B subunit and an echinococcus granulosus EgM123 protein; the active ingredient of the EgM123 protein recombinant mycobacterium smegmatis vaccine can express the recombinant mycobacterium smegmatis of the echinococcus granulosus EgM123 protein. The vaccine provided by the invention can stimulate the immune response in a host, improves the immune level of the host, enhances the intestinal mucosal immune response, and plays an important protective role in resisting infection of echinococcosis.

The invention discloses an echinococcus granulosus developmental-stage secretory protein expression gene chip. The echinococcus granulosus developmental-stage secretory protein expression gene chip is characterized in that sequences of isolated cDNA expression sequence labels of echinococcus granulosus protoscolex, vesicle, oncosphere and adult developmental stage secretory proteins are combined with vectors; and the cDNA expression sequence labels are shown in the formulas of SEQ ID No.1 to SEQ ID No.30. The echinococcus granulosus developmental-stage secretory protein expression gene chip has commercial values and scientific research values. Through the novel cDNA expression sequence labels, a genetic map is drawn. The echinococcus granulosus developmental-stage secretory protein expression gene chip has an important effect of research of secretory protein gene expression. The expression sequence labels can realize screening of targets of a cystic echinococcosis-resistant drug and through the expression sequence labels, the optimal echinococcosis-resistant drug molecules are selected out. The echinococcus granulosus developmental-stage secretory protein expression gene chip can be used for screening of a cystic echinococcosis diagnostic marker and can be used for research and development of an echinococcosis diagnostic kit having high specificity. The echinococcus granulosus developmental-stage secretory protein expression gene chip can be used for screening of cystic echinococcosis protective antigens and can be used for research and development of echinococcosis vaccines suitable for the animal husbandry and pet dogs.

The invention relates to the technical field of application of a harmine derivative, and particularly discloses the application of a harmine derivative to preparation of drugs for treating cystic echinococcosis for the first time. In-vitro pharmacodynamic experiment data prove that the harmine derivative can reduce the survival rate of echinococcosis granulosis cyst remarkably; In-vivo pharmacodynamic experiment data prove that the efficacy of the harmine derivative and the efficacy of albendazole are similar in treating cystic echinococcosis; in-vitro-and-vivo safety evaluation experiment data prove that the toxicity of the harmine derivative is lower than that of a harmine monomer with cystic echinococcosis resisting activity. Therefore, a new approach is provided for treating cystic echinococcosis.

The invention discloses a PCR-RFLP detection kit for authenticating and differentiating the infections of echinococcus multilocularis (Em) and echinococcosis shiquicus (Es). The kit comprises an Em and Es universal primer shown in SEQ ID NO. 1 and 2, restriction endonuclease EcoR I and / or Ssp I, a PCR reagent, and echinococcus multilocularis and echinococcosis shiquicus DNA template reference substances. The purpose of authenticating and differentiating the infections of echinococcus multilocularis and echinococcosis shiquicus can be achieved by extracting the DNA of the single cyst of the sample to be detected as a template, carrying out PCR amplification by using the universal primer, purifying a PCR product, carrying out enzyme digestion by using EcoR I or Ssp I incision enzyme, and then carrying out electrophoresis on an enzyme digestion product. According to the kit disclosed by the invention, the time consumption and money consumption due to sequencing, and the cumbersome process of data processing are omitted, a complex operation system is not required, and infection of echinococcus multilocularis or echinococcosis shiquicus to wild rodents can be rapidly, conveniently and economically differentiated and authenticated.

The invention relates to the technical field of spectral recognition, and provides an echinococcosis patient serum spectral recognition method based on principal component analysis and a BP neural network. The method is implemented according to the steps that 1, serum of at least 20 healthy persons and serum of at least 20 echinococcosis patients are sucked separately and placed in a raman spectrometer for full-wavelength scanning and data collecting; 2, collected data is subjected to normalization processing; 3, the data obtained after normalization is subjected to principle component analysis, and scores of all principle components of which the accumulated contribution rates reach 80% are taken as input layer nodes of the BP neural network. According to the method, an echinococcosis spectral diagnosis technical scheme with the high accuracy is established by adopting the method of combining principle component analysis (PCA) with the BP neural network, the diagnosis accuracy rate is high, and operation and implementation are convenient.

The invention belongs to the field of animal disease diagnosis, and concretely relates to a cerebral echinococcosis marker nad5 gene, and a detection method and a kit of Taenia multiceps. A Taenia multiceps imago sample is collected in a laboratory, DNA is extracted from dog night soil, and PCR amplification is carried out by using parts of the sequence of the mitochondrial nad5 gene as the target DNA fragment of Taenia multiceps. A result shows that the length of the amplified target fragment of Taenia multiceps infected dog night soil is consistent with the length of an expected fragment, and is 433bp. A sequencing identification result shows that the homology of the Cerebral echinococcosis marker nad5 gene to parts of the sequence of the nad5 gene of a known Taenia multiceps strain is 99.5%. The nad5 gene is highly specific and sensitive. The nad5 gene can be used as a Cerebral echinococcosis marker. The detection method of Taenia multiceps can rapidly and effectively detect Taenia multiceps eggs in dog night soil, also has high specificity and sensitivity, and can be used for the epidemiology investigation of the Taenia multiceps.

The invention relates to application of an inhibitor in preparation of drugs for treating cystic echinococcosis, and particularly discloses application of an inhibitor for preparing drugs for treatingcystic echinococcosis. The inhibitor is selected from the following inhibitors: a deubiquitinating enzyme inhibitor, an EGFR inhibitor or a combination thereof. The inhibitor can effectively kill echinococcus granulosus (particularly echinococcus granulosus larva), and then the cystic echinococcosis is treated.

The invention discloses application of tauroursodeoxycholic acid salt used as an active ingredient in preparation of a drug for treating echinococcosis. The tauroursodeoxycholic acid salt is used as the active ingredient for killing pathogen-echinococcus granulosus of the echinococcosis; an obvious echinococcosis protoscolex restraining and killing effect is achieved, and thus, the treatment of the echinococcosis is realized.

The embodiment of the invention discloses a bovine echinococcosis granulosa time-resolved fluorescence immunochromatographic assay test strip and a preparation method thereof, and belongs to the technical field of biological assay. A combination pad of the test strip is coated with a goat anti-bovine IgG antibody marked by time-resolved fluorescent microspheres. The nitrocellulose membrane is provided with a detection line (6) and a quality control line (7) which are separated from each other, the detection line is coated with a recombinant multi-epitope antigen, the quality control line is coated with a rabbit anti-goat IgG secondary antibody, and the recombinant multi-epitope antigen is formed by splicing seven peptide fragments (named as Eg-H1). The test strip disclosed by the inventionhas the advantages of simplicity and convenience in operation and good sensitivity and accuracy; overcomes the defects of incapability of field operation and low accuracy of ELISA, colloidal gold andother diagnosis methods, also has good specificity, provides a rapid diagnosis method for epidemiological investigation of bovine echinococcosis granulosa and diagnosis of the disease, and has wide application prospects.

The invention relates to the technical field of an MAPK signal pathway inhibitor, in particular to application of the MAPK signal pathway inhibitor to preparation of drugs for treating cystic echinococcosis. The invention discloses for the first time the application of the MAPK signal pathway inhibitor to preparation of drugs for treating cystic echinococcosis, and further discloses for the first time application of Sorafenib Tosylate Tablets, U0126 or PD184352 in the MAPK signal pathway inhibitor to preparation of drugs for treating cystic echinococcosis. Pharmacodynamic experimental data show that the treating effect of Sorafenib Tosylate Tablets, U0126 or PD184352 on cystic echinococcosis is superior or equivalent to that of clinically commonly used albendazole on cystic echinococcosis, that is, the MAPK signal pathway inhibitor can serve as a novel anti-echinococcosis drug for cystic echinococcosis, especially Sorafenib Tosylate Tablets, U0126 or PD184352, which can serve as a novel anti-echinococcosis drug, so that a novel approach is provided for cystic echinococcosis treatment.

The invention provides an echinococcus granulosus immunogenic protein EG95 constitutive expression vector and a pichia pastoris genetically engineered bacterium containing the expression vector. The applicant successfully realizes secretory expression of EG95 protein in a pichia pastoris constitutive expression system, the concentration of the protein expressed by a triangular flask shaking tablereaches 0.5g / L, and the purity of the target protein reaches 90% or above. Westernblotting tests prove that the expressed recombinant protein has good reactogenicity and specificity (no reaction withother recombinant protein antibodies) with existing sheep echinococcosis EG95 subunit vaccine immune positive serum. The engineered bacterium capable of efficiently and stably secreting and expressingthe EG95 recombinant protein is screened out from constructed p-GAPZ alpha A-EG95 recombinant plasmid transformed pichia pastoris SMD1168, no inducer is needed, the production period is remarkably shortened, the production cost is remarkably reduced, and the production complexity is remarkably reduced.

The invention discloses a polypeptide for preparing an echinococcosis granulosa diagnostic reagent and application thereof. The polypeptide for preparing the echinococcosis granulosa diagnostic reagent is eB2-3 and / or eB3-3, eB2-3 is the polypeptide with the amino acid sequence shown in sequence 1 in a sequence table, and eB3-3 is the polypeptide with the amino acid sequence shown in sequence 2 inthe sequence table An antibody detection kit prepared by taking an eB2-3 conjugate and / or eB3-3 conjugate obtained by coupling the polypeptide and carrier protein as a coating antigen is high in sensibility, high in accuracy and easy and rapid to operate, and the kit is applicable to mass screening detection of grass-roots veterinary departments at all levels and Entry-Exit Inspection and Quarantine Bureau on echinococcus granulosus infection serum antibodies.

The invention discloses application of lithocholic acid serving as an active ingredient in preparation of echinococcosis granulosis cyst treatment medicine. The echinococcosis granulosis cyst treatment medicine can kill pathogeny-echinococcosis granulosis cyst tapeworms of echinococcosis granulosis cyst through lithocholic acid, has the remarkable restraining and killing effects, and can be used for treating echinococcosis granulosis cyst.

The invention discloses an ELISA (Enzyme-linked Immuno Sorbent Assay) antigen detection kit of animal echinococcosis granulosa. The detection kit is prepared from an anti-EgAgB8 / 2 protein monoclonal antibody pre-coated ELISA plate, a sample diluting solution, a positive control solution, a negative control solution, an enzyme conjugate, a washing solution, a substrate color developing solution anda stopping solution. An anti-EgAgB8 / 2 protein monoclonal antibody is prepared through the following method: firstly, amplifying EgAgB8 / 2 gene cDNA (complementary Deoxyribonucleic Acid) by utilizing RT-PCR (Reverse Transcription-Polymerase Chain Reaction) and cloning an EgAgB8 / 2 gene; constructing a prokaryotic expression vector pET / EgAgB8 / 2 and expressing in escherichia coli BL21(DE3); inducing the expression of the prokaryotic expression vector by utilizing IPTG (isopropyl-beta-d-thiogalactoside); centrifuging and collecting thalli; after dissolving with a bacterium lysis solution and carrying out ultrasonic crushing; centrifuging and collecting supernatant; purifying by utilizing Ni-NTA glue; immunizing a BALB / c mouse by utilizing the purified EgAgB8 / 2 protein, so as to obtain a hybridoma cell line capable of stably secreting the anti-EgAgB8 / 2 protein monoclonal antibody; selecting the excellent monoclonal antibody to produce an antibody and purifying. The kit disclosed by the invention can be used for qualitative detection of an animal echinococcosis granulosa antigen and has the advantages of stable antibody source, good specificity, high sensitivity and good repeatability.

The invention discloses a mobile trackable system for echinococcosis detection. The system comprises a client and a server, wherein the client comprises an obtaining module and an output module; the server comprises a selection module, a sampling module, a calculation module, a comparison module, a result judgement module and a storage module; the system furthermore comprises a cloud database which is capable of storing and managing diagnosis information of diagnosed persons; and a display module comprises a search unit and a search result display unit, and is capable of searching corresponding reaction area images and result information according to unique code information of the diagnosed persons, so as to trace recognition results. The mobile trackable system for echinococcosis detection only needs to obtain an original image which comprises a reaction area and a code image on an echinococcosis IgG immunodetection kit test strip, so as to process and recognize the original image, users do not need to read the result, and the system directly outputs the recognition result, so that the problem of mistaken adjustment caused by the reason that the recognition result reading requireshigh personal professionality and the recognition result is easy to be manually judged by naked eyes is solved. The client of the system can be conventional convenient-to-carry equipment such as mobile phones, tablet computers and the like, so that the detection process is not limited by territoriality and timeliness.

The invention discloses an antigen for detecting a sheep echinococcosis antibody and a preparation method of an agar diffusion plate, and belongs to the technical field of biological detection. The preparation method of the antigen comprises the following steps: performing enzyme digestion connection on a sheep echinococcosis EG95 protein nucleic acid sequence, and constructing the sheep echinococcosis EG95 protein nucleic acid sequence on a plasmid vector; then performing converting into escherichia coli, constructing an expression strain, performing IPTG induced expression, and performing crushing to obtain an inclusion body; and performing washing, cracking, centrifuging, purifying, dialyzing, concentrating, freeze-drying, and diluting with PBS. The preparation method of the agar diffusion plate comprises the following steps: adding water and PBS into agarose, performing heating, adding polyethylene glycol 6000, MES and sodium chloride, performing cooling, performing punching, and sealing the bottom. The antigen and the agar diffusion plate provided by the invention have the advantages of high sensitivity, strong specificity and strong universality, have good biosafety, are easyfor large-scale production, are especially suitable for clinical detection of sheep serum by grassroots veterinarians, and have wide popularization and application value.

The invention discloses a gene engineering vaccine resistant to sheep echinococcosis infection. The vaccine is a protein rEg.P29 vaccine which is prepared by extracting RNA from a China strain of Echinococcus granulosus, obtaining Eg.P29 gene molecules through RT-PCR, building a P29 / pET28a prokaryotic expression vector and performing IPTG induced expression and recombination in Escherichia coli BL21. The vaccine is collected under CCTCCM2015425. Immunoprotection verification of an Echinococcus granulosus recombinant vaccine is performed on sheep, and immune protection efficiency reaches 94.8%. Vaccine candidate molecules are expected to be applied to preparing vaccines resistant to sheep echinococcosis infection and expected to play a great role in preventing and treating liver echinococcosis.

The invention discloses a specific detection antigen of bovine echinococcosis granulosa and application of the specific detection antigen. Specific detection antigen protein of the bovine echinococcosis granulosa is (a1) or (a2): (a1) protein of an amino acid sequence shown as SEQ ID NO.1, and (a2) protein obtained by carrying out substitution and / or deletion and / or addition of one or more amino acid residues on an amino acid sequence of the protein defined by the (a1) and with the same function. Time-resolved fluorescent microspheres are used as tracers, immunochromatographic test strips areprepared, the clinical operation is simple, the diagnosis is rapid, the sensitivity is high, and the specificity is high.

A new preventing and treating technology of sweet potato ditylenchus destructor thorne disease is characterized by including the steps of reinforcing potato seed seedling quarantine, selecting an anti-disease species, reasonably conducting crop rotation, eliminating disease residual bodies, conducting chemical preventing and treating and the like. The problem that the ditylenchus destructor thornedisease exists in the existing sweet potato planting process is solved. The yield per mu of sweet potatoes is greatly improved by about 40%, and the technology is particularly suitable for planting in mountainous areas.

The invention discloses a novel application of osthole in preparation of an echinococcosis treating drug. The novel application of the osthole in the preparation of the echinococcosis treating drug has the benefits that the application effect of the osthole on the echinococcosis resistance is disclosed for the first time, the application of the osthole is expanded, and one novel echinococcosis treating drug is developed. A cytotoxicity test, an in-vitro echinococcosis protoscolices killing test, an in-vivo hydatid tissue mass growth inhibition test and a drug in-vivo liver and kidney injury test research prove that the osthole has remarkable echinococcosis protoscolices killing effect, can remarkably inhibit growth of hydatid tissue, does not have obvious cytotoxicity when the osthole is in effective concentration and has no influence on liver and kidney functions of a mouse when the osthole is in the effective concentration.

The invention relates to the field of immunocolloidal gold chromatography analysis, in particular to a kit for rapidly detecting natural infection antibodies of animal hydatid. The kit can specifically detect the natural infection antibodies of the hydatid, and has no cross reaction with the vaccine immune antibody. The kit comprises a colloidal gold test paper card and a sample diluent, wherein the colloidal gold test paper card comprises a closed box body and a reaction test strip tiling in the box body, the box body is respectively provided with an observation window and a sample adding hole connected to the reaction test strip; and the sample diluent is a mixed solution including 0.01M PH7.2 PBS and 1% tween-80. The invention provides a simple, rapid and scientific method for diagnosing echinococcosis infection, which can quickly detect whether a blood sample contains natural infection antibodies of animal hydatid.

The invention relates to the technical field of cuminal pharmaceutical application, in particular to an application of cuminal in preparation of a medicine for treating echinococcosis granulosa. The invention discloses that cuminal has an inhibiting effect on echinococcus granulosus protoscolex for the first time, cuminal with different concentrations has certain inhibiting and killing effects onechinococcus granulosus protoscolex, and a new medicine is provided for treating echinococcus granulosus.

The invention relates to a visual saliency and SIFT characteristic based echinococcosis protoscolex survival rate detection method, and belongs to the technical field of detection methods. According to the visual saliency and SIFT characteristic based identification method, calculation is carried out on part of an image, only a suspected protoscolex area obtained from a saliency map is taken intoconsideration, a lot of time can be saved in the aspect of SIFT characteristic extraction, a k-means algorithm is used to cluster SIFT characteristic vectors of a sample image to change high-dimensioncharacteristic description of the SIFT characteristic vectors, the problems including too complex calculation and too much time consumption are solved, the calculating efficiency of target searchingvia the SIFT characteristics is improved, the SIFT characteristics in the salient area are more stable, and the problems that manual counting causes a manual error, low working frequency, long counting time and high workload are solved, and the correct rate of identification is ensured.

The invention discloses Tibetan medicine for treating echinococcosis.The Tibetan medicine is prepared from active ingredients or active ingredients and pharmaceutically acceptable excipients, and the active ingredients are prepared from ambrettle seed, Semen cassiae and frankincense according to a certain weight ratio.The Tibetan medicine can be prepared into any common oral dosage forms, has efficacy of diminishing inflammation and stopping pain and mainly functions in treating grasserie, rheumatism, rheumatoid, lepriasis, stroke, echinococcosis, skin diseases, swelling disease and prostatic swelling.

The invention discloses an image processing device, method and equipment, ultrasonic equipment, an ultrasonic system and a readable storage medium, and the device comprises: an obtaining module whichis used for obtaining a to-be-processed ultrasonic image of the liver; the segmentation and classification module that is used for carrying out classification and segmentation on the to-be-processed liver ultrasonic image to obtain a segmentation image and a target classification result corresponding to the to-be-processed liver ultrasonic image, wherein the target classification result comprisesa lesion disease classification result or a liver lesion-free result, and the lesion disease classification result comprises a liver echinococcosis disease classification result or a non-liver echinococcosis disease classification result; and when the target classification result is a disease classification result of a focus, the segmented image comprises a liver contour segmentation line and a focus contour segmentation line. According to the invention, the segmentation and classification module is utilized to identify lesions of liver echinococcosis and other liver space occupying disease types, so that the misdiagnosis rate and the missed diagnosis rate are reduced; and the liver contour is segmented in the segmented image, so that the accuracy of the focus contour is improved.