15 results about "Clonorchis sinensis" patented technology

The invention discloses a preparation method of a double-enzyme inorganic functional nanoflower and application of the double-enzyme inorganic functional nanoflower in preparation of a competitive ELISA (enzyme-linked immuno sorbent assay) kit for detecting clonorchiasis sinensis. Belongs to the technical field of clonorchis sinensis detection. The invention provides a preparation method of a double-enzyme-inorganic functional nanoflower composite material, which comprises the following steps: reacting clonorchis sinensis monoclonal antibody, HRP (horse radish peroxidase) and copper phosphate solution at room temperature for 12-48 hours, and coprecipitating to obtain the functional nanoflower composite material. By utilizing the characteristic that the HNFs can catalyze a substrate TMB, species limitation caused by a traditional enzyme-labeled secondary antibody is broken, the detection sensitivity is improved, and multi-host detection of clonorchis sinensis is realized.

The invention belongs to the technical field of parasite detection, and particularly relates to a detection method of clonorchis sinensis for non-disease diagnosis purpose, which comprises the following steps: taking a primer group A for targeted amplification of clonorchis sinensis genes as a target primer, and taking a primer group B for targeted amplification of escherichia coli genes as an internal reference primer. Two sets of primer groups are combined, the primer group B for amplifying escherichia coli genes is used as an internal reference primer, whether nucleic acid of a sample and a quality control product is successfully extracted or not and whether a detection result is credible or not in the detection process can be monitored, and the technical problem that a conventional clonorchis sinensis detection technology cannot monitor whether an amplification experiment is abnormal or not is solved.

The invention provides a novel ecological feed capable of preventing multiple grass-bream diseases, the novel ecological feed comprises a feed matrix and an ecological feed leavening agent, and the ecological feed leavening agent comprises probiotics with surface expression antigens, the antigen is any one or more of a grass carp reovirus antigen, a flavobacterium johnsonii antigen, an aeromonas veronii antigen and a clonorchis sinensis antigen. The novel ecological feed with a disease-resistant function in a parabramis pekinensis breeding mode is prepared for the first time, the resistance of the parabramis pekinensis to various pathogens can be enhanced, and the survival rate of the parabramis pekinensis in the disease attack peak period can be increased by 50% or above after immunization. An oral immunization mode is adopted and can be completed in the feeding process, and the immunization and breeding cost is greatly saved.

The present invention discloses a method for preparing a dual-enzyme inorganic functional nanoflower and its application in preparing a competitive ELISA kit for detecting Clonorchis sinensis. This invention belongs to the technical field of Clonorchis sinensis detection. The present invention provides a method for preparing a dual-enzyme inorganic functional nanoflower composite material. The composite material is prepared by reacting a Clonorchis sinensis monoclonal antibody, HRP, and a copper phosphate solution at room temperature for 12-48 hours, followed by coprecipitation to obtain the functional nanoflower composite material. By utilizing the ability of HNFs to catalyze the substrate TMB, the method overcomes the species restrictions imposed by traditional enzyme-labeled secondary antibodies, improves detection sensitivity, and enables multi-host detection of Clonorchis sinensis.

The invention discloses a clonorchis sinensis recombinant protease inhibitor Cystatin-2 and a polarization effect of the clonorchis sinensis recombinant protease inhibitor Cystatin-2 on macrophages. The protease inhibitor Cystatin-2 is derived from clonorchis sinensis, can remarkably promote polarization of macrophages, and is an immunomodulatory molecule with application prospects.

The invention discloses an application of clonorchis sinensis ubiquitin-like protein NEDD8 as a clonorchis sinensis infection diagnosis target, and relates to the technical field of biological detection, and the amino acid sequence of the ubiquitin-like protein NEDD8 is as shown in SEQ ID NO: 1. According to the invention, key insect source molecules related to infection immunity are screened, the coding gene Csnedd8 of ubiquitin-like protein NEDD8 is successfully cloned from cDNA of clonorchis sinensis for the first time, efficient inducible expression is realized in Escherichia coli BL21 (DE3) by constructing pET302 / NT-His recombinant plasmids, and the high-purity recombinant protein NEDD8 is obtained through purification. ELISA detection results prove that the NEDD8 can be specifically recognized by serum of infected animals (rats and rabbits) and clinical patients, has low reactivity with serum of healthy people, especially shows a higher detection rate for early infected patients, and solves the core technical problem of early diagnosis target deletion. The colloidal gold immunochromatography test strip developed on the basis of the recombinant protein realizes rapid and on-site diagnosis of clonorchiasis sinensis, and is simple and convenient to operate and suitable for popularization.

The invention relates to the discovery of the virus consisting of Opisthorhabdovirus (ORV), Opisthorphenuilivirus (OPLV), Opisthorsivirus (OSV),Clonorsivirus (CSV), Clonorhabdovirus (CRV), Clonorphenuilivirus (CPLV), Schistohaemavirus (ShV), Schistohaematogalivirus (ShTLV), Schistohaemendornavirus (ShMV) and Schistorhabovirus (ShRV) from the Class I carcinogen trematodes Schistosoma haematobium, Opisthorchis viverrini and Clonorchis sinensis and provide evidence of fluke-borne infection of parasitized vertebrate hosts.. The invention provides viruses, nucleic acids, proteins, peptides, and antibodies useful for diagnosis, treatment, and prevention of viral infection and associated diseases.

The application provides a specific primer pair and kit for identifying Clonorchis sinensis adult, cysticercus and worm eggs and application, and belongs to the technical field of biological molecule detection. In order to solve the problems of insufficient specificity and insufficient sensitivity of the primer in the basic diagnosis of Clonorchis sinensis, the application discloses a specific primer pair for identifying Clonorchis sinensis adult, cysticercus and worm eggs, wherein the upstream primer sequence of the primer pair is shown as SEQ ID NO. 1, and the downstream primer sequence is shown as SEQ ID NO. 2. The detection method is simple, efficient and objective, the specific primer is used, and the Clonorchis sinensis can be quickly identified through simple PCR technology; the prepared kit is low in cost, high in accuracy and simple in operation, and can meet the operation of non-professionals.

This invention belongs to the field of biomedical technology, specifically relating to microRNA derived from extracellular vesicles of Clonorchis sinensis and its application in Clonorchis sinensis-induced cholangiocarcinoma. This invention extracts CsEVs from the supernatant of adult Clonorchis sinensis and sequences them, ultimately selecting Csi-miR-190-3p as the research subject. It is confirmed that Csi-miR-190-3p can inhibit the migration and invasion of RBE cells in vitro; its target gene is CXCL5, and this inhibitory effect is alleviated after co-transfection with the CXCL5 plasmid, indicating that Csi-miR-190-3p derived from Clonorchis sinensis extracellular vesicles holds promise for providing a new direction for the preparation of drugs to treat Clonorchis sinensis-induced cholangiocarcinoma.

The invention belongs to the technical field of medicines, and particularly relates to an application of Csi-miR-125a derived from clonorchis sinensis extracellular vesicles in cholangiocarcinoma. According to the invention, the CsEVs (extracellular vesicles) of the clonorchis sinensis are successfully extracted from the adult supernatant of the clonorchis sinensis, and the Csi-miR-125a which plays a key role in the pathopoiesis related process of the clonorchis sinensis is screened by sequencing. It is further proved that Csi-miR-125a can significantly promote proliferation of RBE cells (bile duct cancer cells) in an in-vitro environment, and a target gene for promoting proliferation of the RBE cells is BAK1; and when the Csi-miR-125a and the BAK1 plasmid are co-transfected, the promotion effect of the Csi-miR-125a on the proliferation of the RBE cells can be obviously relieved. The result shows that the Csi-miR-125a derived from the clonorchis sinensis extracellular vesicles is expected to provide a brand new direction for preparing the medicine for treating the clonorchis sinensis induced cholangiocarcinoma.

This invention belongs to the field of parasite detection technology, specifically relating to a lyophilized PCR kit for detecting Clonorchis sinensis in fecal samples. The lyophilized PCR kit of this invention includes a lyophilized reaction powder, a positive control, a negative control, and a reconstitution solvent. The lyophilized reaction powder is prepared by freeze-drying a reaction solution containing the following components: (a) a first primer pair and a first probe for detecting the Clonorchis sinensis COX1 gene; (b) a second primer pair and a second probe for detecting the human genomic actin (ACT) gene; (c) PCR reaction components: including hot-start Taq DNA polymerase, dNTPs, and a buffer system; and (d) a lyophilization protectant. This invention targets the Clonorchis sinensis-specific COX1 gene and uses PCR amplification technology to detect fecal samples, accurately identifying Clonorchis sinensis nucleic acid and effectively avoiding cross-reactions with other parasites and intestinal flora. The detection specificity and sensitivity are significantly improved, and the repeatability and stability are excellent.

The application discloses application of a Slug inhibitor in preparation of a therapeutic drug for cholangiocarcinoma induced by Clonorchis sinensis extracellular vesicles, and also includes application in preparation of an inhibitor for epithelial-mesenchymal transition, migration, proliferation and invasion of cholangiocarcinoma cells induced by Clonorchis sinensis.The application further provides a drug for treating cholangiocarcinoma induced by Clonorchis sinensis, which comprises siRNA with Slug gene as a target.The application firstly proves that Clonorchis sinensis extracellular vesicles promote proliferation, migration, invasion and epithelial-mesenchymal transition of cholangiocarcinoma cells, and knocking down Slug in the cholangiocarcinoma cells can inhibit the progress of the physiological activities, thereby providing a new technical option for clinical treatment of cholangiocarcinoma.

The application belongs to the technical field of biological medicine, and particularly relates to application of Csi-miR-31 in regulation of METTL3 / m6A to promote cholangiocarcinoma and M2 polarization. Csi-miR-31-5p is screened as a research object by extracting CsEVs from Clonorchis sinensis adult supernatant and then sequencing; the experimental results show that overexpression of Csi-miR-31-5p can significantly promote proliferation of cholangiocarcinoma cells in vitro, significantly enhance migration and invasion ability of the cholangiocarcinoma cells, inhibit apoptosis of the cholangiocarcinoma cells, induce polarization of macrophages to M2 type immunosuppressive phenotype and reshape tumor immunosuppressive microenvironment. Therefore, Csi-miR-31-5p provided by the application can be used as a high-specificity diagnostic marker and an independent treatment target and a combined immunotherapy target, and provides a new direction and experimental basis for development of precise diagnosis and treatment drugs of cholangiocarcinoma induced by Clonorchis sinensis.

The invention discloses an RPA-PfAgo detection system for clonorchis sinensis and a preparation method of the RPA-PfAgo detection system, and a visual RPA-PfAgo detection method is established on the basis of a conservative COX-1 gene design primer and gDNA of the clonorchis sinensis. By optimizing amplification conditions, the efficiency and the specificity of the RPA reaction are remarkably improved. In order to simplify the operation process and prevent pollution, purification is not needed after the RPA reaction is finished, the product can be directly added into a PfAgo system to react for 25 minutes, the result is interpreted through blue light irradiation, complex instruments and equipment are not needed, and the result can be observed by naked eyes in a short time; the method has the characteristics that the detection method is simple, the multifunctionality is greatly enhanced, and professional personnel operation is not needed.

The present invention relates to the field of detection of intestinal parasites from patients, food or environmental samples, preferably from fecal samples. In particular, the present invention provides a polymerase chain reaction (PCR) based assay for detecting intestinal parasitic infections, in particular infections with parasitic species selected from the group consisting of: Hymenolepis nana, Hymenolepis diminuta, Fasciolopsis buski, Encephalitozoon spp (such as E. intestinalis, E. cuniculi and E. hellem), Enterocytozoon bieneusi, Enterobius vermicularis, Diphyllobothrium latum, Diphyllobothrium nihonkaiense, Schistosoma mansoni mansoni, Blastocystis hominis, Ancylostoma duodenale and liver worms (such as Clonorchis sinensis, Opisthorchis spp. and Metorchis spp.). The present invention further provides materials for the methods of the present invention, such as primers, primer pairs and probes. Preferably, the methods of the present invention are multiplex real-time fluorescence quantitative PCR assays for rapid determination of clinically important intestinal parasites.