65 results about "Echinococcoses" patented technology

The invention belongs to the field of bioengineering and provides an Echinococcus granulosusglutathione transferase gene a base sequence of which is shown in SEQ ID NO: 1. The invention also provides a protein encoded by the Echinococcus granulosusglutathione transferase gene. The amino acid sequence of the protein encoded by the Echinococcus granulosusglutathione transferase gene is shown in SEQ ID NO: 2, or is the same as 1st-219th amino acid sequences as shown in SEQ ID NO: 3. The recombinant protein can be used for immunodiagnosis of cystic echinococcosis patients and can be used for coating a plate; and the serums of the patients having cystic echinococcosis, alveolitoid echinococcosis, cysticercosis and other parasite diseases and healthy people are detected by using an enzyme-linked immunosorbent assay (ELISA), and the obtained sensitivity and specificity are respectively 72.41% and 92.36%.

The invention discloses a PCR (Polymerase Chain Reaction) detection kit for cystic echinococcosis of a dog. According to the method, a pair of primers is designed according to a 12S rDNA (ribosome Deoxyribose Nucleic Acid) sequence conserved region of pathogene echinococcus granulosus of the cystic echinococcosis, a PCR reaction is carried out with the DNA contained in the wastes of an animal to be detected serving as a template, and then a comparison is carried out relative to an attached negative control and a positive control, in order to determine whether the detected animal infects the echinococcosis. The detection kit is simple and easy to use, and high in sensitivity and stability and can be used for monitoring the health condition of a pet dog.

The invention discloses an immunochromatography test strip for detecting cystic echinococcosis and a preparation method thereof. The test strip comprises a hemofiltration membrane sample pad, a gold-marking pad, a cellulose membrane and a water absorption pad, wherein the gold-marking pad is tightly connected with the hemofiltration membrane sample pad and comprises an antibody labeling colloid gold probe of anti-echinococcus antigen advantage special antibody subtypes, the cellulose membrane is tightly connected with the gold-marking pad, the water absorption pad is tightly connected with the other end of the cellulose membrane, a detection line and a quality control line are arranged on the cellulose membrane, the detection line comprises crude antigens aiming at the cystic echinococcosis, and the quality control line comprises anti-antibodies capable of realizing special combination with the antibodies of the anti-echinococcus antigen advantage special antibody subtypes. The immunochromatography test strip of the invention has the advantages of simplicity, convenience, sensitivity, specificity and high speed, and is applicable to clinical and field use.

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 relates to an anti-echinococcosis antigen monoclonal antibody hybridoma cell strain, an anti-echinococcosis antigen monoclonal antibody and application of the anti-echinococcosis antigenmonoclonal antibody. The anti-echinococcosis antigen monoclonal antibody hybridoma cell strain is classified and named as a hybridoma cell strain XJ10D8D10. The strain is preserved in the China Center for Type Culture Collection (CCTCC) on September 25th, 2017. The preservation address is Wuhan University, Wuhan, China, and the preservation number is CCTCC NO: C201789. According to the invention,an echinococcosis adult surface membrane antigen is extracted; a Balb/c mouse is immunized; splenic lymphocytes secreting anti-echinococcosis adult surface membrane antigen are obtained; the hybridoma cell strain secreting the anti-echinococcosis adult surface film antigen is obtained by fusing the splenic lymphocytes with SP2/0 cells, so that an echinococcosis-resistant adult surface film antigen antibody is obtained, and the echinococcus granulosus infection of dogs can be sensitively detected through fecal antigen detecting by using the antibody as a diagnostic reagent. And necessary conditions are provided for preparation of large-scale general survey detecting kits.

The invention discloses a detection card for quickly quantitative detection of echinococcosis antibody in serum. The detection card includes a detection card case and a test paper strip assembled therein. The test paper strip includes a plastic base board provided with pressure-sensitive adhesive, wherein a sample pad, a marker pad, a nitrocellulose membrane and water absorption paper are successively adhered to the base board. The marker pad is composed of a carrier base layer and a marker. The marker is a film formed by spray-coating the carrier base layer with lanthanum-series fluorescencedetection microspheres and lanthanum-series fluorescence quality control microspheres. The nitrocellulose membrane is coated with an echinococcosis recombinant antigen to form a detection line and rabbit-anti-chicken IgY antibody to form a quality control line. The marker includes the fluorescence detection microspheres labeled by the echinococcosis recombinant antigen and the fluorescence qualitycontrol microspheres labeled by the chicken IgY antibody. The detection card can achieve quick quantitative detection of the echinococcosis antibody in situ and has great practical and application values.

The invention discloses a cystic echinococcosis diagnostic kit and application thereof. The kit comprises a coating antigen composed of an eB2-3 conjugate and/or an eB3-3 conjugate; the eB2-3 conjugate represents a complete antigen obtained by coupling of eB2-3 with carrier protein; the eB3-3 conjugate represents a complete antigen obtained by coupling of eB2-3 with carrier protein; eB2-3 represents polypeptide shown by the sequence 1; and eVP1-2 represents polypeptide shown by the sequence 2. The kit is high in specificity, sensitivity and accuracy, simple and rapid in operation, and suitable for rapidly screening and detecting a lot of serum antibody infected by echinococcus granulosus by veterinary departments and entry-exit inspection and quarantine bureaus in all levels.

The invention discloses a sheep echinococcosis ELISA antibody detection kit and a preparation method and application thereof. In the detection kit, an elisa plate is pre-coated with an echinococcus granulosus cocktail antigen; the echinococcus granulosus cocktail antigen is composed of an EPC1 recombinant antigen, an EgAgB1 recombinant antigen, an EgAgB2 recombinant antigen and an EgAgB4 recombinant antigen. The echinococcus granulosus cocktail antigen is composed of an EPC1 recombinant antigen, an EgAgB1 recombinant antigen, an EgAgB2 recombinant antigen and an EgAgB4 recombinant antigen. Thedetection kit provided by the invention can be used for rapidly detecting the echinococcosis antibody in the sheep serum, is simple to operate, short in consumed time, high in sensitivity and good inspecificity, and is convenient for simultaneously detecting a large number of samples. The sheep echinococcosis ELISA antibody detection kit provided by the invention has important application valuein echinococcosis antibody level detection and epidemiological investigation.

The invention discloses an echinococcus granulosus recombinant protein, which is characterized in that four EgG1Y162-2 protein fragments are connected in series through a linker sequence 'GSGGSG' to form the recombinant protein; experiments show that the recombinant protein vaccine can promote maturation of dendritic cells. The action principle and effect of the recombinant vaccine EgG1Y162-2 (4) are disclosed for the first time, and a foundation is laid for preparing vaccines or diagnostic kits for preventing and treating echinococcosis of people or livestock.

The invention discloses a novel echinococcosis antigen Murinoglobulin-2 protein. The amino acid sequence of the Murinoglobulin-2 protein provided by the invention contains all or part of the following sites: the 31st site to the 39th site, the 89th site to the 96th site, the 107th site to the 115th site, the 179th site to the 200th site, the 203rd site to the 216th site, the 225th site to the 242nd site, the 255th site to the 273rd site, the 282nd site to the 300th site, the 309th site to the 315th site, the 336th site to the 354th site and the 379th site to the 390th site of SEQ ID No.1. The Murinoglobulin-2 protein provided by the invention can be used for clinically detecting the human echinococcosis. The Murinoglobulin-2 protein is used for detecting plasma of 14 cases of postoperative echinococcosis people and plasma of 6 cases of normal Tibetan people, the positive detection rate is 86%, and the negative detection rate is 100%. According to the invention, a hydatid antigen library is expanded, and a certain contribution is made to diagnosis of human-derived echinococcosis.

The invention provides an echinococcosis granulosa gene engineering vaccine. The sequence of the diagnostic antigen P-29 gene of the echinococcosis granulosa Uruguay strain is retrieved from the Genbank of www.ncbi.nlm.nih.Gov, the P-29 gene is amplified through RT-PCR, with the total RNA of the echinococcosis granulosa pathogens of echinococcosis patients in Ningxia region, China as the template, and the DNA sequence of the amplified gene is consistent with that of the gene of the Uruguay strain. The cloned P-29 gene and the expression vector pET-28a are recombined through the recombinant DNA technology to transform and construct the engineered escherichia coli strain. The P-29 recombinant protein is prepared and the obtained P-29 recombinant protein immune mice are attacked by the protoscolex pathogens of echinococcosis patients in Ningxia region. Through computation in the experiments, the protective immunity of EgP-29 is 96.6%. The P-29 recombinant protein is proved to have immunology effect of the vaccines.

The embodiment of the invention provides a method for detecting cystic echinococcosis. The method comprises the following steps: coating an ELISA plate with an Ag5 antigen which is recombined and induced by a prokaryotic expression system and is purified in vitro; diluting serum of a subject in proportion, and adding the diluted serum and known negative and positive serum into the plate holes of the ELISAplate for reaction; and measuring the OD450 value of each plate hole, and calculating the sample value of each plate hole according to the publicity. Clinical serum sample detection shows that the detection specificity and sensitivity are higher than those of a commercial kit, and false positive results are effectively eliminated. The indirect ELISA method established in the invention provides a more accurate and effective detection means for serological diagnosis of echinococcosis.

The invention relates to the technical field of preparation of medicines for treating echinococcosis, and relates to a composition of a photodynamic combination drug and a preparation method and an application thereof. The composition comprises a photosensitizer or/and a chemical drug; wherein the photosensitizer is one or more of chlorin e6, viltipofen, indocyanine green, porphin sodium, 5-aminolevulinic acid, temopofen and talapofen, and the chemical drug is one or more of albendazole, albendazole sulfoxide, mebendazole, flubendazole, oxfendazole, artesunate, peganum harmala, tetrandrine andsophora japonica. According to the invention, through photodynamic therapy, the permeability of the hydatid and the hydatid cyst wall is increased; according to the invention, the target accumulationof chemotherapeutic drugs in the echinococcosis and the cyst wall is realized, the synergistic treatment effect is achieved, the resistance of the echinococcosis and the cyst to external drugs is reduced, the curative effect of the anti-echinococcosis drugs is improved, the administration dosage and the toxic and side effects of the drugs can be greatly reduced, and the application prospect in the preparation of echinococcosis treatment drugs is excellent.

The invention relates to the technical field of preparation of medicine for treating echinococcosis, in particular to a composition combining DNA damage causing compounds with DNA damage repair inhibitors, and a preparation method and application of the composition. The composition comprises the DNA damage causing compounds and the DNA damage repair inhibitors. The DNA damage causing compounds areone or more kind of materials of harmine, harmine derivatives, adriamycin, dactinomycin, daunorubicin, etoposide and teniposide; and the DNA damage repair inhibitors are one or more kind of materialsof an RAD51 inhibitor and a BRCA1 inhibitor. Through combined use of the DNA damage causing compounds and the DNA damage repair inhibitors, the composition provided by the invention is used for treating the echinococcosis; the effect of realizing the damage to worm body DNA by the DNA damage causing compounds and realizing the inhibition on the DNA damage repair by the DNA damage repair inhibitors at the same time can be achieved; the worm body DNA damage repair process is completely blocked; finally, the worm body apoptosis is caused; the anti-echinococcosis effect is achieved; and the anti-echinococcosis treatment effect is improved.

The invention discloses an early diagnosis and treatment biomarker for alveolar echinococcosis and application thereof. The invention protects application of a substance for detecting target proteinsin preparation of a product for diagnosis or auxiliary diagnosis of alveolar echinococcosis. The target proteins include (a1) or (a2) or (a3): the (a1) comprises ALDH1A1 protein, TAGLN2 protein and FLN alpha protein; the (a2) comprises any two of ALDH1A1 protein, TAGLN2 protein and FLN alpha protein; and the (a3) comprises ALDH1A1 protein or TAGLN2 protein or FLN alpha protein. The inventors of the invention use proteome correlation analysis of liver and plasma to detect potential candidate biomarkers in AE patients. Results show that ALDH1A1, TAGLN2 and FLN alpha may be early prediction indexes of the AE patients and can be used as candidate diagnosis indexes of early AE patients.

The invention discloses a neoantigen Cystatin protein for echinococcosis and a preparation method of the neoantigen Cystatin protein. The amino acid sequence of the Cystatin protein provided by the invention contains all or part of the following components: the 1st to 25th sites, the 32nd to 56th sites, the 64th to 84th sites, the 89th to 95th sites, the 141st to 147th sites and the 155th to 168th sites of SEQ ID No.1. The neoantigen Cystatin protein is found through a GeLS-MS/MS technology, and the neoantigen Cystatin protein for echinococcosis can be used for manufacturing an ELISA kit and used for clinically detecting human echinococcosis. The Cystatin protein is used for detecting plasma of 14 cases of postoperative echinococcosis people and plasma of 6 cases of normal Tibetan people, the positive detection rate is 93%, and the negative detection rate is 100%. According to the invention, a hydatid antigen library is expanded, and a certain contribution is made to diagnosis of human-derived echinococcosis.

The invention discloses a free DNA sequence derived from echinococcus granulosus. The length of the free DNA sequence is equal to 107 basic groups and is represented by SEQ ID NO.24. Besides, the invention further discloses a screening method of the free DNA sequence derived from the echinococcus granulosus. The screening method comprises the following steps: (S1) acquiring a large number of freeDNA sequences from a blood sample of a patient with cytic echinococcosis by virtue of a re-sequencing technique; (S2) carrying out comparative analysis by virtue of a bioinformatics method so as to screen out echinococcus granulosus-sourced free DNA sequences; and (S3) carrying out verification and screening in patients with the echinococcosis and healthy people through a fluorescence quantitativePCR method, so as to obtain the free DNA sequence derived from the echinococcus granulosus. Besides, the invention further discloses application of the free DNA sequence derived from the echinococcusgranulosus in preparation of products for diagnosing and detecting the echinococcosis. A detection result shows that the free DNA sequence has relatively high sensitivity and specificity in detectionof the patient with the echinococcosis and has excellent application prospects in the diagnosis and detection fields of the echinococcosis.