29 results about "Echinococcus granulosus" patented technology

The invention relates to the technical field of tapeworm identification, and provides a kit for identifying five target tapeworms by using specific primers and application of the kit. All mainstream genotypes of genes of Echinococcus granulosus, Echinococcus multilocularis, Taenia vesicula, Taenia multiceps and Taenia canis are compared, and a primer pair capable of universally detecting nucleic acids of five tapeworms is obtained by taking conserved regions of the genes as diagnostic targets. The rapid detection of the five pathogens is realized by combining with a digoxin-labeled nucleic acid lateral chromatography test paper. A product obtained by isothermal amplification is detected by a nucleic acid detection test strip and then is directly observed by eyes, and the result is interpreted within 15-20 minutes, so that the operation is simple and convenient. The method provided by the invention has wide application prospects in the aspects of large-scale screening and detection of whether dogs are infected with echinococcus granulosus, echinococcus multilocularis, taenia vesicula, taenia multiceps and tapestoma dogs or not in pasturing areas, especially in areas with epidemic echinococcosis, and evaluation of the insect expelling effect of the dogs fed with praziquantel on the tapeworms.

A hypothetical protein gene of Echinococcus granulosus and its encoded protein application belong to the field of immunology technology. To address the current stagnation in the development of vaccines for canine anti-Echinococcus granulosus infection and the lagging screening of protective antigens, the present invention first constructs a cDNA expression library of three-day-old Echinococcus granulosus larvae. The library is then immunologically screened using positive serum from dogs infected with Echinococcus granulosus. Through second-generation sequencing and data analysis, an antigenic protein of three-day-old Echinococcus granulosus larvae is screened from the library. The amino acid sequence of the antigenic protein is shown in SEQ ID NO.2, and the nucleotide sequence of the encoding gene is shown in SEQ ID NO.1. It has been verified that the antigenic protein can specifically react with positive serum from dogs infected with Echinococcus granulosus, and has high application value in the development of canine anti-Echinococcus granulosus vaccines and diagnostic reagents.

The invention relates to a gene marker group, a primer, a kit and application of the gene marker group, the primer and the kit in detection of echinococcosis, and belongs to the technical field of biology. Wherein the gene marker group comprises at least one of (c1) to (c230). The gene marker group can be used for designing multiple detection primers for detecting echinococcus.

The invention relates to the technical field of molecular biology, and discloses a primer probe set for species identification of echinococcus granulosus, and the primer probe set comprises a first primer probe set for targeting echinococcus granulosus, a second primer probe set for targeting echinococcus multilocularis and / or a third primer probe set for targeting echinococcus stone. According to the primer probe group, synchronous, rapid and high-specificity identification and detection of three kinds of echinococcus, namely echinococcus granulosus (Eg), echinococcus multilocularis (Em) and echinococcus stone (Es), are realized. When the primer probe group is used for identifying and detecting the three kinds of echinococcus, the detection sensitivity reaches 200 copies / mL, and the positive coincidence rate reaches 100%. The primer probe group realizes synchronous and accurate identification of echinococcus species, and has a wide application prospect in molecular epidemiological investigation and port quarantine.

The invention discloses an echinococcus granulosus PLGA-EgG1Y162 nano subunit vaccine and a preparation method thereof, and relates to the field of biomedical engineering and parasitic disease prevention and treatment. According to the method, echinococcus granulosus EgG1Y162 recombinant protein (the amino acid sequence is as shown in SEQ ID NO.1) is taken as an antigen core, and is stably coated in a polylactic acid-glycolic acid copolymer (PLGA) nano carrier through an innovative embedding process, so that the nano subunit vaccine in a brand new form is constructed. Compared with pure EgG1Y162 recombinant protein, the nanoparticles prepared by the preparation method disclosed by the invention not only have innovated components, but also obviously improve the slow release performance of drugs, and can realize precise enrichment and long-acting release of antigens. The invention fills up the blank of research on the echinococcus granulosus PLGA nano subunit vaccine, provides a novel technical scheme for prevention and treatment of echinococcosis, and has important application value and market prospect in the field of biological medicine.

A hypothetical protein gene of Echinococcus granulosus and its encoded protein application belong to the field of immunology technology. To address the current stagnation in the development of vaccines for canine anti-Echinococcus granulosus infection and the lagging screening of protective antigens, the present invention first constructs a cDNA expression library of three-day-old Echinococcus granulosus larvae. The library is then immunologically screened using positive serum from dogs infected with Echinococcus granulosus. Through second-generation sequencing and data analysis, an antigenic protein of three-day-old Echinococcus granulosus larvae is screened from the library. The amino acid sequence of the antigenic protein is shown in SEQ ID NO.2, and the nucleotide sequence of the encoding gene is shown in SEQ ID NO.1. It has been verified that the antigenic protein can specifically react with positive serum from dogs infected with Echinococcus granulosus, and has high application value in the development of canine anti-Echinococcus granulosus vaccines and diagnostic reagents.

The invention discloses a rapid detection method for echinococcus granulosus based on RPA-CRISPR / Cas, and belongs to the technical field of molecular diagnosis. The method comprises the following steps: extracting DNA of a sample to be detected; the specific primers are used for RPA isothermal amplification, a reaction system contains a buffer solution, upstream and downstream primers, RPA enzyme, magnesium acetate and template DNA, and reaction is carried out at 39 DEG C for 20 minutes; and mixing the amplification product with LbCas12a protein, sgRNA and a fluorescence report probe, carrying out CRISPR / Cas reaction at 37 DEG C for 30 minutes, and judging the result through a real-time fluorescence signal. Constant-temperature rapid amplification is achieved through RPA, high-sensitivity and high-specificity detection of the echinococcus granulosus can be achieved under the site condition by combining high-specificity recognition and trans-cleavage activity of CRISPR / Cas12a, the sensitivity reaches the single-site copy number, operation is easy and convenient, complex temperature control equipment is not needed, and the method is suitable for primary medical treatment and site quarantine.

The invention relates to a molecular marker primer, a kit and a method for rapidly identifying an Echinococcus granulosus gene, and belongs to the technical field of biomolecular marker detection. The molecular marker primer for rapidly identifying the Echinococcus granulosus gene comprises a forward primer EG PrimerF, a reverse primer EG PrimerF, a reverse primer EG PrimerF, a reverse primer EG PrimerF, a reverse primer EG PrimerF, a reverse primer EG PrimerF and a reverse primer EG PrimerF, the sequence of the reverse primer EG Primer R is GAATGCTGC TGACTAACGAG, and the sequence of the reverse primer EG Primer R is GAATGCTGC TGACTAACGAG. According to the molecular marker primer, the kit and the method for rapidly identifying the Echinococcus granulosus gene, the target DNA is subjected to PCR amplification by utilizing the primer, a specific band near 480bp is detected through agarose gel electrophoresis, and the Echinococcus granulosus gene can be rapidly and accurately identified. Based on existing genome data and a traditional molecular biological technology, the problem that a traditional detection method is insufficient in specificity is solved, and an efficient scheme is provided for early diagnosis and large-scale screening of echinococcosis.

The invention relates to a preparation process of an artemisinin-metal organic framework composite bone hydatid resistant sustained release microsphere, which comprises the four steps of material preparation, artemisinin-metal organic framework compound preparation, sustained release microsphere preparation and post-treatment and characterization, the preparation process of the artemisinin-metal organic framework composite bone hydatid resistant sustained release microsphere comprises the following steps: selecting a UI0-66 material; according to the artemisinin emulsion, the structure can be kept intact for 28 days under the physiological condition, TPGS is adopted as a compound stabilizer, the metal organic framework is pretreated, and TPGS is added into a water phase and an oil phase in the emulsification stage, so that the molecular diffusion coefficient of artemisinin is reduced to 1 / 5, the binding capacity of the metal organic framework and artemisinin is improved by 2.3-2.5 times, the cell intake is increased, and the stability of the artemisinin emulsion is improved. The TPGS is preferentially explained in an enzymolysis environment, a time-dependent release channel is formed, and the stability problem of the artemisinin is solved.

This invention discloses a bivalent subunit vaccine against Echinococcus granulosus and Echinococcus cantonensis, and its preparation method. The bivalent subunit vaccine comprises the immunogen dEG95 / dEC95 antigen protein and a pharmaceutically acceptable carrier. The amino acid sequence of the dEG95 protein is shown in SEQ ID No. 1, and the amino acid sequence of the dEC95 protein is shown in SEQ ID No. 2. Currently, there are no vaccines that simultaneously protect against Echinococcus granulosus and Echinococcus cantonensis infection. This invention uses the dEG95-dEC95 heterodimeric protein as the antigen, and the prepared bivalent subunit vaccine against both Echinococcus granulosus and Echinococcus cantonensis has low production cost and a simple production process. It can simultaneously prevent infection by both Echinococcus granulosus and Echinococcus cantonensis, and has many advantages such as safety, high efficiency, low cost, and high antigen purity.

The invention discloses a novel anti-echinococcus monoclonal antibody, a hybridoma cell strain and application of the novel anti-echinococcus monoclonal antibody, and belongs to the technical field of biomedicine and immunology. The preservation number of the hybridoma cell strain Anti-Em-11 disclosed by the invention is CGMCC (China General Microbiological Culture Collection Center) No. 46598. According to the present invention, the specific monoclonal antibody is successfully prepared and identified, and Western blot and indirect immunofluorescence experiments prove that the obtained monoclonal antibody has high specificity on the recombinant protein. The invention lays a solid foundation for serological diagnosis of the echinococcosis and remarkably enriches the existing serological detection technical system.

According to the echinococcus granulosus vaccine based on the self-assembly peptide, a peptide fragment, rich in dominant epitopes, in EgG1Y162-2 is connected to a Q11 amino terminal through '-SGSG-', due to the self-assembly effect of the Q11 peptide, the peptide fragment rich in dominant epitopes is repeatedly expressed on the surface of nanofibers, and therefore EgG1Y162 dominant multi-epitope self-assembly nano-polypeptide is formed; the EgG1Y162 dominant multi-epitope self-assembled nano polypeptide is adopted in the vaccine, so that the immune recognition and response of a body are effectively enhanced.

The present invention relates to a classification model for echinococcosis granulosus and brucellosis in sheep based on infrared spectroscopy, a method for establishing the same, and a detection device. A method for establishing a classification model for echinococcosis granulosus and brucellosis in sheep based on infrared spectroscopy includes: (1) collecting fresh blood from healthy and diseased sheep, extracting serum to obtain serum samples; (2) detecting the serum samples using a Fourier transform infrared spectroscopy (FT-IR) instrument to obtain FT-IR spectral data; (3) performing processing such as baseline correction and normalization on the FT-IR spectral data to obtain processed spectral data; (4) after reducing the dimensionality of the processed spectral data using the principal component analysis method (PCA), establishing a linear discriminant analysis (LDA) or support vector machine (SVM) classification model to obtain a classification model for echinococcosis granulosus and brucellosis in sheep. The classification model established by the present invention based on machine learning algorithms and FT-IR spectroscopy has the potential for rapid screening of sheep with echinococcosis granulosus and sheep with brucellosis.

An Echinococcus granulosus antigen Eg73 and its encoding gene and application belong to the field of immunology technology. The present invention utilizes a three-day-old Echinococcus granulosus larvae cDNA expression library to screen the Eg73 antigen protein in a high-throughput, unbiased manner, and further provides a method for preparing the antigen protein and its application, aiming to provide a new antigen target for the development of candidate antigens for canine anti-Echinococcus granulosus vaccines. This solves the current problem of stagnant vaccine development for canine anti-Echinococcus granulosus infection and lagging protective antigen screening. The amino acid sequence of the antigen protein is shown in SEQ ID NO.6, and the nucleotide sequence encoding the antigen protein is shown in SEQ ID NO.7. It has been verified that the antigen protein can specifically react with positive serum from dogs infected with Echinococcus granulosus, and has high application value in the research and development of canine anti-Echinococcus granulosus vaccines and diagnostic reagents.

The application discloses an in-vitro RNA interference method for a Taenia solium lactate dehydrogenase gene, related reagents, a preparation method and uses. The reagent for reducing expression of the Taenia solium lactate dehydrogenase gene can specifically interfere with an EmLDH-B gene, and provides a new idea for in-depth research on a Taenia solium infection mechanism and development of a potential treatment target.

This invention discloses a multiplex PCR primer combination, kit, and detection method for detecting Echinococcus granulosus. The multiplex PCR primer combination includes a second primer combination and a third primer combination: the second primer combination consists of upstream and downstream primers specific to Echinococcus multilocularis, with sequences shown in SEQ ID NO. 3 and SEQ ID NO. 4, respectively; the third primer combination consists of upstream and downstream primers specific to Echinococcus shiquensis, with sequences shown in SEQ ID NO. 5 and SEQ ID NO. 6, respectively. This invention saves more time than conventional PCR, is more cost-effective than quantitative real-time PCR, and has lower requirements for operators and equipment, showing great promise for the detection and control of echinococcosis.

The invention discloses a construction method and application of a mouse hepatic echinococcosis granulosis cyst model, and relates to the technical field of biology. The construction method comprises the following steps: under the guidance of B ultrasound, carrying out liver envelope injection on a mouse, injecting an echinococcus granulosus protoscolex into liver parenchyma through envelope puncture, and carrying out feeding and cystic focus screening to obtain the mouse liver echinococcus granulosus model. According to the invention, a percutaneous liver membrane injection method under the guidance of B ultrasound is adopted for the first time, and an injection site is accurately positioned under the monitoring of a real-time image, so that the colonization rate of an original head section in the liver is remarkably improved, and meanwhile, the risk of operative trauma and abdominal cavity dissemination is avoided. The method disclosed by the invention is simple and convenient to operate, low in injury, short in time consumption, high in success rate, controllable in focus position and size and good in repeatability. Besides, according to the method, mechanical injuries to the liver and other visceral organs are greatly reduced, the inflammatory reaction and rejection risks are reduced, and a more reliable animal model is provided for pathogenesis research, drug screening and vaccine evaluation of the echinococcosis granulosa.

An Echinococcus granulosus antigen Eg142, its encoding gene, and its application belong to the field of immunology technology. The present invention utilizes the Eg142 antigen protein, obtained through high-throughput, unbiased screening of a three-day-old Echinococcus granulosus larvae cDNA expression library, to develop a new antigen candidate for a canine Echinococcus granulosus vaccine. This solves the current problems of stagnation in the development of canine Echinococcus granulosus vaccines and the lag in screening for protective antigens. The amino acid sequence of the antigen protein is shown in SEQ ID NO.3, and the nucleotide sequence encoding the Eg142 protein is shown in SEQ ID NO.4. It has been verified that the antigen protein can specifically react with positive serum from dogs infected with Echinococcus granulosus, and has high application value in the development of canine Echinococcus granulosus vaccines.

The application discloses a SPON1-1 protein fragment and application thereof in human echinococcosis. The polypeptide comprises one or more of the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6. The recombinant protein comprises the amino acid sequence shown in SEQ ID NO:7; the mutated amino acid sequence has at least 80%, 85%, 90%, 95%, 98%, 99% identity with the amino acid sequence of the recombinant protein, and maintains the function of the recombinant protein. The application further discloses application of the polypeptide or the recombinant protein or a mutation thereof or the marker combination in preparation of an antibody against echinococcosis or a diagnostic agent for diseases caused by echinococcosis granularis. The new antigen SPON1-1 of echinococcosis can be used to prepare an ELISA kit and is used for clinical detection of human echinococcosis; and can be purified in a large amount, so that large-scale screening of human echinococcosis is facilitated.

The application discloses a UP protein fragment and application thereof in human echinococcosis. The polypeptide comprises one or more of the amino acid sequences as shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9. The application further discloses application of the polypeptide or the marker combination in preparation of an antibody against echinococcosis or a diagnostic agent for diagnosing a disease caused by echinococcus granulosus. The new antigen UP of echinococcus in the application can be used to prepare an ELISA kit and used for clinical detection of human echinococcosis, and can be purified in a large amount, thereby facilitating large-scale screening of human echinococcosis.

The invention belongs to the field of biology, and discloses an echinococcus granulosus mRNA vaccine and a preparation method thereof. After codon optimization is carried out on an EgG1Y162 protein coding sequence of echinococcus granulosus, a T7 promoter, a 5 '-untranslated region (5' UTR) sequence and a Kozak sequence are added to the front end of the sequence, a 3 '-untranslated region (3' UTR) and a poly A tail (Poly A) are added to the rear end of the sequence, an mRNA vaccine template DNA sequence is formed, and then mRNA is obtained, and the mRNA has the advantages of being good in immunogenicity, high in protection strength and the like when existing in an echinococcus granulosus vaccine form.

The invention discloses a duplex mRNA (messenger ribonucleic acid) vaccine for echinococcus granulosus and coenuriasis and a preparation method of the duplex mRNA vaccine, and the duplex mRNA vaccine comprises mRNA for simultaneously expressing Eg95 recombinant protein and TM16 recombinant protein and LNP for encapsulating the mRNA, and the LNP is recorded as Eg95-TM16 mRNA-LNP. According to the invention, Eg95 and TM16 recombinant proteins and Eg95-TM16 mRNA-LNP are respectively used for immunizing mice and sheep, and results show that the Eg95 and TM16 mixed recombinant proteins and the Eg95-TM16 mRNA-LNP can activate immune response of CD8 + T lymphocytes so as to play a protective role in resisting parasitic infection; meanwhile, the Eg95-TM16 mRNA-LNP shows good stability and consistent immunogenicity in a sheep body and mainly causes humoral immune response which mainly takes IgG antibody level rise, and antibody dynamics is similar to mouse data. The Eg95-TM16 mRNA-LNP immune group enhanced cell factor induction is superior to the immune stimulation capability of recombinant protein, so that the Eg95-TM16 mRNA-LNP can simultaneously play the immune protection role of the echinococcus granulosus and the coenuriasis, and the Eg95-TM16 mRNA-LNP is expected to become a novel immunoprophylaxis means for resisting the echinococcus granulosus and the coenuriasis.

The invention provides an echinococcus granulosus antibody, a kit for detecting echinococcus granulosus pathogen from excrement and application of the echinococcus granulosus antibody. Firstly, a monoclonal antibody 4E6 is provided, the monoclonal antibody 4E6 can stably and efficiently recognize soluble antigens of adult echinococcus granulosus, soluble antigens excreted and secreted by the echinococcus granulosus and soluble antigens of eggs of the echinococcus granulosus, and the monoclonal antibody 4E6 has high titer and good specificity on the various mixed antigens. Therefore, the antibody can be used for preparing a kit for detecting echinococcus granulosus eggs in canine animal excrement or environment. The preparation method of the monoclonal antibody 4E6 for resisting echinococcus granulosus provided by the invention is simple, the antibody purification process is simple, the efficiency is high, and the cost is low. The monoclonal antibody 4E6 provided by the invention is used for detecting polluted echinococcus granulosus in excrement of canine animals or environment, and has the advantages of rapidness, sensitivity, high specificity, simplicity in operation, low cost and the like.

The application discloses a preparation and application of a fine granular echinococcus and a Canadian echinococcus EG95 / EC95 protein combination; the recombinant protein combination comprises dEG95 and dEC95 amino acid sequences; the dEG95 amino acid sequence is a modified amino acid sequence obtained by deleting a signal peptide region at an N terminal and a transmembrane region at a C terminal of EG95 of narrow fine granular echinococcus; the EG95 gene sequence is shown as SEQ ID NO. 1, and the amino acid sequence is shown as SEQ ID NO. 2; the dEC95 amino acid sequence is a modified amino acid sequence obtained by deleting a signal peptide region at an N terminal and a transmembrane region at a C terminal of EC95 of Canadian echinococcus; the EC95 gene sequence is shown as SEQ ID NO. 4, and the amino acid sequence is shown as SEQ ID NO. 5. The method has the advantages of high protein expression amount, simple process, low production cost and the like.

The invention discloses a primer combination, a kit and a detection method for simultaneously detecting three echinococcosis pathogens, and belongs to the technical field of parasitic pathogen detection. The primer combination comprises a primer sequence as shown in SEQ ID NO. 1-6. The primer combination comprises a primer sequence as shown in SEQ ID NO. The primer pair combination designed by the invention does not interfere with each other when multiple PCR amplification is carried out in the same system under the same condition, is strong in specificity and high in sensitivity, and is designed for echinococcosis pathogens (echinococcus multilocularis, narrow-sense echinococcus granulosus and echinococcus stone). On the basis, a multiplex PCR method capable of identifying narrow-sense echinococcus granulosus, echinococcus multilocularis and echinococcus stone in faeces and tissues is established, the method is high in specificity and sensitivity, has the advantages of high flux, simplicity, convenience and rapidness, can remarkably improve the working efficiency, reduces the time and economic cost, and is suitable for popularization and application. The method has a wide application prospect in the field of echinococcosis molecular epidemiological investigation.

The invention provides a novel positive antigen of echinococcus granulosus and application of the positive antigen. Specifically, the invention provides an application of Echinococcus granulosus lactic dehydrogenase (EgLDH) or a gene thereof or a detection reagent thereof, and the EgLDH or the gene thereof or the detection reagent thereof is (i) used for preparing a positive antigen of the Echinococcus granulosus (or echinococcosis granulosus), and (ii) used for preparing a negative antigen of the Echinococcus granulosus (or echinococcosis granulosus), and (ii) used for preparing a negative antigen of the Echinococcus granulosus (or echinococcosis granulosus (or echinococcosis granulosus (or echinococcosis granulosus). And / or (ii) is used for preparing a reagent or a kit for diagnosing echinococcus granulosus (or echinococcus granulosus). The antigen disclosed by the invention has very high diagnostic sensitivity and specificity on echinococcus granulosus, and the diagnostic efficiency is far higher than that of other conventional echinococcus granulosus antigens.

The invention belongs to the technical field of genetic engineering, and relates to a subunit vaccine for preventing and treating canine echinococcosis and application. The vaccine is a recombinant fusion protein vaccine for preventing canine echinococcosis granulosus tapeworm infection, novel recombinant fusion protein based on canine echinococcosis EG95 antigen is constructed and expressed, and is prepared into a subunit vaccine for immunizing a dog, and the canine echinococcosis can be effectively prevented by stimulating cellular immunity and humoral immunity in the dog. Therefore, the life transmission cycle of echinococcus granulosus tapeworms is blocked, and a preventive vaccine is provided for echinococcosis terminal host prevention and control.

The present invention relates to the field of biotechnology, and in particular to primer combinations, reagent combinations, kits, and their applications and methods in identifying Echinococcus granulosus G1 type and Echinococcus multilocularis. The present invention is based on the principle of recombinase polymerase amplification combined with lateral flow chromatography test strips, designs specific primers for the mitochondrial genomes of Echinococcus granulosus G1 type and Echinococcus multilocularis, and achieves accurate identification of Echinococcus granulosus G1 type and Echinococcus multilocularis by optimizing the concentrations of relevant reagents and reaction conditions. The present invention has high sensitivity, strong specificity, high accuracy, and good repeatability. In addition, the entire detection process is convenient and fast, requiring only a constant temperature heating instrument and no other expensive instruments. The present invention greatly shortens the detection time of pathogens and reduces the equipment conditions required for detection, thereby meeting the needs of rapid detection of Echinococcus granulosus G1 type and Echinococcus multilocularis on-site or at grassroots units, and has broad application prospects.

The present invention discloses the use of a recombinant protein of a muscle-attached antigen of Echinococcus granulosus in the preparation of a drug for treating asthma. The present invention uses a recombinant protein of a muscle-attached antigen of a strain of Echinococcus granulosus from mainland China (NCBI gene sequence number Z29075) to intervene in an asthma mouse model caused by ovalbumin (OVA). The results show that the muscle-attached antigen recombinant protein of the present invention is used to prepare a drug for treating asthma. The drug can effectively increase the number of Th1 and Treg cells, reduce the number of Th2 and Th17 cells, reduce inflammatory factors in asthma, and increase the IL-10 type cytokine that is reduced in asthma. It can have a good preventive, intervention and / or relief effect on asthma. In addition, the drug also has the characteristics of few side effects and low drug resistance.