40 results about "Trichinella species" patented technology

The present invention relates to paramyosin gene of new trichina antigen, constitution of the vector containing the gene, prokaryon expression vector containing the gene and the expressed protein. The present invention also relates to the application of the gene and its coded protein. When the recombinant protein of trichina paramyosin gene is used in immunizing animal, high titer antibody and effective immunoprotection against attacting infection will generate.

The invention provides a kit for detecting cysticercus cellulosae, swine trichinella and swine toxoplasmosis. The kit comprises sterile deionized water, polymerase chain reaction (PCR) liquid, thermus aquaticus deoxyribonucleic acid (Taq DNA) polymerase, a GoldView DNA dye, bromophenol blue sample loading buffer solution, a standard substance and a reference substance. In the detection kit provided by the invention, DNA of the cysticercus cellulosae, the swine trichinella and the swine toxoplasmosis in a specimen to be detected can be detected quickly and accurately only by one reaction, and the kit can also be used for the molecular epidemiological survey and curative effect monitoring of cysticercus cellulosae, swine trichinella and swine toxoplasmosis infection. The method has simple preparation steps and is low in cost, easy to operate, time-saving and labor-saving, a large number of reagents and consumables are saved, and the work efficiency is improved. The method is high in detection sensibility, specificity and accuracy, and the false positive rate is reduced.

The invention discloses a food-borne verminosis fast enzyme immunity detection method and a perforated detection kit, wherein, an enzyme immunity detection reagent comprises an antigen coating plate A, enzyme conjugate liquid B, cleaning solution C, a substrate D, a color developing reagent E, sample diluent F, terminator G, a positive reference substance H and a negative reference substance I; the perforated detection kit is used during the detection process which comprises sample dilution, sample adding reaction, enzyme adding reaction and color developing reaction; the antigen coating plate A adopts quick coating technology, and antigens are coated on a general purpose polystyrene mipor plate in advance; two polyethylene glycols with molecular weight Mr of 20000 and 6000 are added into the enzyme conjugate liquid B. The invention solves the problems of the prior detection method like long operating time, trivial operation, instable reagents, incomplete match and so on, and has the advantages of quick speed, convenient operation, high reliability, good detection results and so on. The invention is used for enzyme immunity detection of cysticercosis cutis, paragonimiasis, distomatosis and trichinosis.

The invention provides a trichina and tumor cell related protein gene. The preparation method of protein gene comprises the following steps of: determining the related antigen existing between a tumor cell antigen and trichina positive serum and between a trichina antigen and tumor cell positive serum by utilizing an enzyme-linked immunosorbent assay; and screening a trichina cDNA (complementary deoxyribonucleic acid) expression library by utilizing SP2 / 0 hyperimmuneserum to obtain the related protein gene. The trichina and tumor cell related protein gene is a novel protein gene substance; tumor cells are sensitive to a variant identification substance, and the trichina and tumor cell related protein gene can produce strong specific and nonspecific immunoreaction to tumors, and the trichina and tumor cell related antigen is easy for preparation and industrialization.

The invention provides a fluorescence immunochromatography detection test paper strip of trichinellosis as well as a preparation method and application thereof and belongs to the technical field of detection of fluorescence immunodetection. In order to more rapidly, stably and accurately detect trichinization, the invention provides a fluorescence immunochromatography detection test paper strip oftrichinosis. The fluorescence immunochromatography detection test paper strip of trichinosis comprises a sample pad, a combination pad, a chromatography membrane, a water absorption pad and a substrate, wherein time-resolved fluorescence microsphere-coupled goat anti-swine IgG is labeled on the combination pad; a detection line and a quality control line are arranged on the chromatography membrane; a trichinella spiralis larvae ES mixed antigen composed of a trichinella spiralis larvae excretion and secretion material ML-ESP antigen and a trichinella intestinal phase spiralis larvae excretionand secretion material iML-ESP antigen is sprayed on the detection line; rabbit anti-goat IgG is sprayed on the quality control line; all the components are combined and prepared into the test paperstrip. The test paper strip can be applied to rapid detection of swine trichinellosis.

The invention discloses a preparation method of a recombinant protein and in particular relates to a TsSerpin recombinant protein and application thereof. The preparation method of the TsSerpin recombinant protein comprises the following steps: carrying out reverse transcription, with total RNA (Ribonucleic Acid) extracted from pig trichina muscle larva as a template and Oligo(dT)18Primer as a primer; with the obtained reverse transcription Cdna (Deoxyribose Nucleic Acid) as a template and SEQ ID No.1 and SEQ ID No.2 as primers, carrying out PCR (Polymerase Chain Reaction) amplification, to obtain pig trichina TsSerpin gene; and carrying out the treatment including recombinant plasmid construction, escherichia coli conversion and the like to obtain target protein. The recombinant protein disclosed by the invention can be used for preparing an antigen for detecting trichinosis.

The invention relates to a preparation method of Rubus delavayi Franch. plant extracts and a pharmaceutical composition thereof as well as application of the Rubus delavayi Franch. plant extracts to anti-trichinella spiralis, and belongs to the field of traditional Chinese medicines and natural medicines. The preparation method of a anti-trichinella spiralis medicine made of the extract as an active ingredient comprises the following steps: taking and drying a whole plant of the Rubus delavayi Franch. plant and pauperizing the whole plant of the Rubus delavayi Franch. plant to reach 20 meshesto 30 meshes; performing extraction by using an organic solvent ( petroleum ether or chloroform or ethyl acetate or acetone or methanol or ethanol) or water. The invention also discloses application of the Rubus delavayi Franch. plant to preparation of an anti-trichinella spiralis agent; 0.01g to 30g of the plant is applied to a substrate or one group, and can be optionally combined with a carrierand / or a medium, so that the plant has higher anti-trichinella spiralis activity. The preparation method of the Rubus delavayi Franch. plant extracts and the pharmaceutical composition thereof as well as application of the Rubus delavayi Franch. plant extracts to the anti-trichinella spiralis disclosed by the invention have the advantages that the adaptability of Rubus delavayi Franch. medicinalmaterials is widened, and a medicinal value of the Rubus delavayi Franch. medicinal materials is improved.

The invention provides a preparation method of a cervical carcinoma cell associated trichina antigen antibody, and also provides medical application of the antibody. Tumor cells are relatively sensitive to a foreign identifier, the cervical carcinoma cell associated trichina antigen can generate strong specific and nonspecific immune reactions on tumor, and simultaneously, the tumor cell associated trichina antigen is easy to prepare and industrialize.

The invention discloses a method for detecting a trichinella circulating antigen by utilizing IgY-McAb sandwich ELISA (enzyme-linked immnuo sorbent assay). With an anti-trichinella muscle larval ES antigen egg yolk antibody IgY as a capture antibody and an anti-trichinella ES antigen McAb as a detection antibody, the preparation method of the IgY comprises the following steps of: adding the trichinella muscle larval into a culture medium to carry out sterile culture, purifying and dialyzing supernate and then concentrating and drying to obtain the ES antiagen, determining protein concentration and then immunizing roman legehenne by utilizing the ES antigen, collecting the IgY from the produced egg yolk by adopting a saturate ammonium sulphate precipitation method, purifying and determining concentration, and sub-packaging for later use; and the preparation method of the McAb comprises the following steps of: establishing a hybridoma cell strain secreting the anti-trichinella muscle larval ES antigen, cloning by adopting a limiting dilution method, purifying McAb by applying an octanoic acid-ammonia sulphate method, determining the concentration and sub-packaging for later use. The invention has the advantage that a new method which has high sensitivity and specificity and has wide application prospect is provided for early detection of trichinella CAg in blood serum of an infected animal.

The invention discloses a fluorescence immunochromatography test strip capable of detecting early infection of pig trichina and a production method thereof, and belongs to the technical field of serological detection. In order to detect trichina infection more quickly, conveniently and sensitively, the test strip comprises a sample pad, a combination pad, a chromatography membrane, a water absorption pad and a bottom plate, and a fluorescent probe is sprayed on the combination pad. The fluorescent probe comprises a time-resolved fluorescent microsphere coupled goat anti-rabbit IgG serving as an indication probe and a time-resolved fluorescent microsphere coupled trichina ES mixed antigen serving as a capture probe, wherein the trichina ES mixed antigen consists of a trichina muscle larva excretion secretion ML-ESP antigen and a trichina intestinal muscle larva excretion secretion iML-ESP antigen; and a detection line and a quality control line are arranged on the chromatography membrane, mouse anti-pig IgG is sprayed on the detection line, and rabbit anti-goat IgG is sprayed on the quality control line. The test strip can be used for on-site rapid detection of trichina.

The invention aims to provide an sHSPs recombinant invasive lactobacillus plantarum live vector DNA vaccine. Lactobacillus is selected as a vector to construct the recombinant invasive lactobacillus plantarum. The lactobacillus plantarum has immunoregulation ability and carries tumor antigen to inhibit tumor growth, and trichina sHSPs as a heteroantigen can break immune tolerance, such that obvious anti-tumor immunity is generated. The lactobacillus plantarum carries the related antigen gene sHSPs so as to provide non-specific immunoregulation ability of the lactobacillus, and the sHSPs can be delivered to a host cell to express the related antigen sHSPs so as to induce a specific immune response. The recombinant lactobacillus plantarum carrying the trichina sHSPs gene exert non-specific immunoregulation ability of the lactobacillus and can also induce specific anti-tumor immunity. Meanwhile, as a probiotic, the lactobacillus has high safety, can be orally taken, and is convenient to use and easy to popularize.

The invention discloses a primer set for detecting trichina gene by loop-mediated constant temperature amplification, an application and a method. The primer set comprises a pair of outer primers F3 / B3, a pair of inner primers FIP / BIP and a pair of loop primers LF / LB, wherein the primer set is used for detecting trichina polypide and ova. The method comprises the steps of extracting sample genomeDNA to be tested, using genome genomic DNA as a formwork, adding the genome DNA to constant temperature amplification reaction liquid, performing a water bath reaction to obtain reaction products, wherein the constant temperature amplification reaction liquid comprises LAMP buffer liquid, a primer set, Bst 2.0 DNA polymerase, pyrophosphatase, dNTP, MgSO4 and ddH2O, and mixing the reaction productswith the detection liquid. If the liquid is changed into blue, results are positive, and if the liquid maintains colorless, the results are negative. The primer set can quickly and accurately detectthe trichina genome DNA, is high in sensibility and high in specificity, has good stability, is simple to operate, can well meet requirements of staff of different levels, and is easy to promote for base.

The invention discloses a competitive monoclonal antibody-based trichinosis antibody detection kit and a preparation method thereof, and belongs to the technical field of fluorescence immunoassay. Theinvention discloses a method for detecting trichina infection more quickly, stably and accurately. The invention provides the competitive monoclonal antibody-based trichinosis antibody detection kit,a test strip comprises a sample pad, a chromatography membrane, a water absorption pad and a bottom plate, the chromatography membrane is provided with a detection line and a quality control line, the detection line is sprayed with a rabbit anti-Ts-WN10 polyclonal antibody, and the quality control line is sprayed with rabbit antimouse IgG; a competitive reagent contains a monoclonal antibody Ts-WN10-1H9 secreted by a hybridoma cell strain WN10-1H9 and a recombinant antigen Ts-WN10; and the microbiological preservation number of the hybridoma cell strain WN10-1H9 is CGMCC No. 18316. The kit prepared by the invention is high in specificity and sensitivity, and can be used for quickly detecting the swine trichinopathy.

The invention belongs to the field of immunobiology, and relates to H-2d restricted Th epitope P2 of trichina paramyosin, its composition and an application thereof. The invention concretely relates to Th2 epitope of trichina paramyosin, which contains an amino acid sequence shown in any one of SEQ ID NO: 2-5. The epitope of the present invention can stimulate / promote significant proliferation of splenic lymphocyte, can stimulate / promote cells increase of mice spleen cell CD4<+>T, can stimulate / promote differentiation of Th0 cell to Th2, can stimulate / promote increase of Th1-type cytokine IFN-gamma, IL-2 and Th2-type cytokine IL-4, IL-5, and can stimulate / promote differentiation of Th0 cell to Th2, after mice is immunized, a Th2 immunization reaction is generated through induction, and the epitope has potential for preparing vaccines such as multi-epitope vaccine.

The invention relates to the field of immunology, and particularly relates to a B cell epitope polypeptide of a serine protease inhibitor (Ts-WM5) with high expression in the larvae phase of trichinamuscle, a hybridoma cell strain, a monoclonal antibody and application thereof. The hybridoma cell strain capable of secreting an anti-Ts-WM5 protein specific antibody is obtained, and the Ts-WM5 protein specific B cell epitope polypeptide recognized by the monoclonal antibody secreted by the hybridoma cell strain is identified, so that the B cell epitope polypeptide has important significance indiagnosis of trichinosis and has important significance in establishing sandwich ELSIA for competitive ELISA and detection of circulating antigens for detection of multiple hosts of trichina and development of subunit vaccines.

Trichinella can be detected in a tissue extract sample. A suitable kit includes (a) a detection carrier containing a first antibody against one or more antigens of Trichinella, and (b) (i) a second antibody against one or more antigens of Trichinella, wherein the second antibody is bound to a signal molecule, or (b) (ii) contains one or more antigens of Trichinella bound to a signal molecule, wherein the antigens of (b) (ii) are configured so as to dissolve binding of the antigens of (a) to the first antibody by competitive displacement.

The invention provides an anti-trichina small heat shock protein (sHSPs) chicken egg yolk antibody and a preparation method thereof, and also discloses a medical application of the egg yolk antibody in tumor resistance. The traditional Chinese medicine composition has the characteristics of enhancing animal immunity, being non-toxic and the like, can be used as an oral preparation medicine for treating tumors, and can be prepared into oral liquid and the like as food for preventing tumors; the egg yolk antibody for poultry such as chicken can be orally taken, is convenient to use, non-toxic, low in cost, safe and convenient for large-scale industrialized production.

The invention provides a recombinant TsPKA-C protein and application thereof and particularly relates to application of the recombinant TsPKA-C protein as taenia solium / cysticercerci detection antigen and application of the recombinant TsPKA-C protein to preparation of a porcine cysticercosis detection kit. The recombinant TsPKA-C protein can specifically detect taenia solium / cysticercerci and has no cross reaction with porcine thin-neck cysticercerci, trichinella sui and the like.

The invention provides a multiple DPO-PCR (dual priming oligonucleotide-polymerase chain reaction) primer combination for detecting trichinella spiralis, toxoplasma gondii and schistosome. The sequences of the primer combination are as shown in SEQ ID NO:1 to SEQ ID NO:6 correspondingly. The invention also provides a multiple DPO-PCR method for detecting the trichinella spiralis, the toxoplasma gondii and the schistosome. By adopting the method, qualitative detection on the muscular tissue or viscera sample of animals suffering from diseases can be realized. Experiments indicate that the detection method provided by the invention has high specificity, high flux and high speed, the detection efficiency is improved, and a more effective method is provided for detecting three animal origin parasitic zoonoses of the trichinella spiralis, the toxoplasma gondii and the schistosome.

The invention discloses a trichina antibody double-hole rapid detection card and a detection method, the trichina antibody double-hole rapid detection card comprises a PVC base plate, a water absorption pad, a nitrocellulose membrane, a marker pad, a sample pad, a filter substance pad and a dilution pad which are installed in a shell, the nitrocellulose membrane is fixed on the PVC base plate, the left end of the nitrocellulose membrane is in lap joint with the water absorption pad, the right end of the nitrocellulose membrane is in lap joint with the dilution pad, a marker pad is arranged at the right position of the middle of the nitrocellulose membrane, a sample pad is arranged on the marker pad, a detection line and a quality control line are arranged between the water absorption pad and the marker pad on the nitrocellulose membrane, and a window hole, a sample adding hole, a cleaning hole and a quantitative marking line are formed in the shell. A sample is added through the middle sample adding hole, and a diluent is added through the rear cleaning hole, so that the sensitivity and the precision of a detected object can be improved, and the characteristics of rapidness, simplicity and convenience of a rapid diagnosis detection method are kept.

The invention belongs to the field of immunobiology, and relates to H-2d restricted Th epitope P2 of trichina paramyosin, its composition and an application thereof. The invention concretely relates to Th2 epitope of trichina paramyosin, which contains an amino acid sequence shown in any one of SEQ ID NO: 2-5. The epitope of the present invention can stimulate / promote significant proliferation of splenic lymphocyte, can stimulate / promote cells increase of mice spleen cell CD4<+>T, can stimulate / promote differentiation of Th0 cell to Th2, can stimulate / promote increase of Th1-type cytokine IFN-gamma, IL-2 and Th2-type cytokine IL-4, IL-5, and can stimulate / promote differentiation of Th0 cell to Th2, after mice is immunized, a Th2 immunization reaction is generated through induction, and the epitope has potential for preparing vaccines such as multi-epitope vaccine.

The invention provides a trichina 7TR protein human single-chain antibody and a preparation method. The trichina 7TR protein human single-chain antibody is obtained through carrying out prokaryotic expression on a trichina 7TR single-chain antibody and carrying out purification and renaturation; screening a Tomlinson human single-chain antibody library; and carrying out inducible expression on a screened positive single-chain antibody secretion strain and purifying by adopting a saturated ammonium sulfate precipitation method and Protein A affinity chromatography. The single-chain antibody can be used for preparation of anti-lung cancer drugs.

The invention provides a kit for detecting cysticercus cellulosae, swine trichinella and swine toxoplasmosis. The kit comprises sterile deionized water, polymerase chain reaction (PCR) liquid, thermus aquaticus deoxyribonucleic acid (Taq DNA) polymerase, a GoldView DNA dye, bromophenol blue sample loading buffer solution, a standard substance and a reference substance. In the detection kit provided by the invention, DNA of the cysticercus cellulosae, the swine trichinella and the swine toxoplasmosis in a specimen to be detected can be detected quickly and accurately only by one reaction, and the kit can also be used for the molecular epidemiological survey and curative effect monitoring of cysticercus cellulosae, swine trichinella and swine toxoplasmosis infection. The method has simple preparation steps and is low in cost, easy to operate, time-saving and labor-saving, a large number of reagents and consumables are saved, and the work efficiency is improved. The method is high in detection sensibility, specificity and accuracy, and the false positive rate is reduced.

The invention provides a hybridoma cell strain, a monoclonal antibody secreted from the hybridoma cell strain and application thereof and belongs to the technical field of prevention and control of trichinella spiralis (T.spiralis). Against the technical problem of specific diagnosis on the T.spiralis, the hybridoma cell strain has a preservation number of CGMCC No.18317. Tests prove that the monoclonal antibody Ts-ZH68-2A40Ab secreted from the hybridoma cell strain can compete against T.spiralis infected positive porcine serum to combine with Ts-ZH68 antigen, and has a recognition peptide being <222>GVDRSATCQGDSGGP236. The monoclonal antibody and the Ts-ZH68 protein B cell epitope peptide recognized by the monoclonal antibody can be used for preparing reagents or vaccines for diagnosing or preventing T.spiralis infection, and lays foundation to establishment of a T.spiralis serological diagnosing method.

The invention provides an application of recombinant trichina thioredoxin peroxidase 2 in preparation of an anti-trichina infection vaccine, which belongs to the technical field of immunoregulation, and the recombinant trichina thioredoxin peroxidase 2 can induce a mouse body to generate Th2 type immune response, induce proliferation of CD4<+> T cells of a mouse and convert macrophages into M2 type cells by the recombinant trichina thioredoxin peroxidase 2. The immune response reactions play a main role in the trichinella spiralis infection resisting process; therefore, the recombinant trichina thioredoxin peroxidase 2 as an antigen molecule can be used for preparing an anti-trichina infection vaccine.

A trichinella spiralis competition ELISA antibody detection kit and a detection method thereof belong to the technical field of serological immunological detection. In order to detect multi-host Trichinella infection more stably and accurately, the invention provides a Trichinella competitive ELISA antibody detection kit, which contains: a 96-well reaction plate whose bottom plate is coated with Trichinella Ts-WN10 recombinant antigen, Biotin-labeled monoclonal antibody Ts-WN10-1H9 solution, horseradish catalase HRP-labeled avidin solution, washing solution, TMB chromogenic substrate, stop solution, and negative quality control; wherein, the negative The quality control is a solution of biotin-labeled monoclonal antibody Ts‑WN10‑1H9 at a concentration of 1 μg / mL. The kit prepared by the invention has the characteristics of strong specificity, high sensitivity, etc., and can be used for antibody detection of multi-host trichinellosis such as pigs / rats / humans.

The invention relates to the field of immunology, in particular to a B cell epitope polypeptide of a trichinella spiralis intestinal stage cysteine ​​protease inhibitor (Ts-WN10), a hybridoma cell line, a monoclonal antibody and applications thereof. The present invention obtains a hybridoma cell line that can secrete anti-Ts-WN10 protein-specific antibody, and identifies the Ts-WN10 protein-specific B cell epitope polypeptide recognized by the monoclonal antibody secreted by it, which is effective against trichinellosis The diagnosis is of great significance, and it is of great significance for the establishment of competitive ELISA and sandwich ELSIA for the detection of circulating antigens for the detection of Trichinella multi-host and the development of subunit vaccines.

Trichinella can be detected in a tissue extract sample. A suitable kit includes (a) a detection carrier containing a first antibody against one or more antigens of Trichinella, and (b) (i) a second antibody against one or more antigens of Trichinella, wherein the second antibody is bound to a signal molecule, or (b) (ii) contains one or more antigens of Trichinella bound to a signal molecule, wherein the antigens of (b) (ii) are configured so as to dissolve binding of the antigens of (a) to the first antibody by competitive displacement.

The invention relates to a trichina gene Ts27 and a protein encoded by the same and also relates to a recombinant vector and a recombinant cell containing the gene Ts27 and an antibody specifically combined with the protein encoded by the gene Ts27. The invention also relates to an application of the gene Ts27, the protein encoded by the gene Ts27 and the antibody to preparation of a detection reagent for diagnosing trichinization of humans and animals, and a detection kit. After the gene Ts27 recombinant protein is immune to animals, an antibody with a high titer can be generated; and the gene Ts27 recombinant protein can be specifically identified by animal or human serums infected by trichinas.

The invention provides application of triclosan in preparation of a medicine for killing parasitic nematodes, and belongs to the technical field of medicinal chemistry. Triclosan has an obvious lethal effect on trichina in all stages, is low in dosage and high in safety, can be used as one of candidate molecules for treating nematode diseases, and can be used for treating nematode diseases. The important reference value is realized on the development of new nematode killing medicines.