24 results about "Cryptosporidium parvum" patented technology

The invention belongs to the technical field of biology, and particularly relates to a cryptosporidium parvum infection immunosuppression mouse model and a construction method thereof. Aiming at the problem that the development of cryptosporidiosis drugs and vaccines is seriously limited due to the fact that no model for successfully infecting mice with cryptosporidium parvum exists at present, the invention provides the construction method of the cryptosporidium parvum infection immunosuppression mouse model, which comprises the following steps: firstly, carrying out gavage treatment on the mice for 4-7 days by adopting cyclophosphamide; constructing to obtain an immunosuppressive mouse; feeding PBS (Phosphate Buffer Solution) containing cryptosporidium parvum oocysts through the mouth, and infecting for 7-21 days; and when oocysts are continuously detected from excrement of the mouse and the content of the oocysts is higher than the infection amount, constructing the cryptosporidium parvum infection immunosuppression mouse model. The cryptosporidium parvum infection immunosuppression mouse model is successfully constructed, can be used for subsequent drug and vaccine development tests for cryptosporidium parvum diseases, and has a good application prospect.

The invention relates to an anti-cryptosporidium micromolecular lead compound of a target cryptosporidium helicase. Comprising the following steps: S1, detecting enzyme activity parameters of a drug target helicase (CpeIF4A) and an inhibition effect of Roc-A on the drug target helicase: expressing and purifying a CpeIF4A recombinant protein containing an MBP tag; detecting the enzyme activity; detecting the inhibition effect of the Roc-A on the enzyme activity; determining the affinity between the Roc-A and the CpeIF4A through a heat transfer experiment; s2, in-vitro drug effect determination: evaluating the direct killing capability of Roc-A on sporozoites; the inhibition effect of Roc-A on cryptosporidium is determined through an in-vitro cell infection model, an EC50 value is obtained, and the intracellular period of the Roc-A mainly playing the inhibition effect is determined; detecting the toxicity of Roc-A to host cells to obtain a TC5 value), and obtaining an in-vitro insect-resistant selectivity index (SI); and S3, in-vivo drug effect test: establishing a C57BL / 6IFN-gamma gene knockout mouse infection model to evaluate the effectiveness and safety of Roc-A. The invention determines that the Roc-A has the potential of being further developed into the cryptosporidium-resistant drug, and verifies that the targeted eIF4A can resist cryptosporidium infection.

The invention belongs to the field of biology, and discloses a cdc42 gene knockout HCT-8 cell line and application thereof, the taxonomic name of the cdc42 gene knockout HCT-8 cell line is Homo sapiens, the cdc42 gene knockout HCT-8 cell line is preserved in Guangdong Microbial Culture Collection Center, and the preservation number is GDMCC NO: 66616; the preservation date is July 01, 2025; the preservation address is the fifth floor of building 59, No.100 courtyard, Xianlie Middle Road, Guangzhou City, Guangdong Province. According to the invention, a cdc42 gene in an HCT-8 cell is knocked out by adopting a CRISPR-Cas9 technology so as to construct a cdc42 gene knocked-out HCT-8 cell line, and the cdc42 gene knocked-out HCT-8 cell line is used as a cryptosporidium parvum cell infection model.

The invention discloses a cryptosporidium parvum AOX gene deleted strain as well as a construction method and application thereof, and the cryptosporidium parvum AOX gene deleted strain disclosed by the invention is obtained by editing an AOX gene of cryptosporidium parvum through a gene editing technology. According to the gene editing technology, the cryptosporidium parvum AOX gene is knocked out by using a CRISPR / Cas9 gene editing technology, so that a cryptosporidium parvum AOX gene deleted strain is obtained; the AOX gene is found to be related to insect toxicity for the first time, and the infection intensity of an AOX gene deleted insect strain in a gamma interferon gene knockout mouse body is remarkably reduced. Research shows that after the AOX gene is deleted from the wild type cryptosporidium parvum IIdA20G1 subtype strain, the strain has the advantage that the toxicity is obviously weakened, and can be used for preparing cryptosporidium parvum attenuated vaccines so as to prevent cryptosporidium parvum infection.

The invention discloses application of a propolis extract in preparation of a medicine for resisting Cryptosporidium parvum infection, and belongs to the technical field of biomedicine.The preparation method of the propolis extract comprises the steps that a propolis raw material is added into an extraction solvent according to the mass-to-volume ratio (g / mL) of 1: (5-20), extraction is conducted for 24-72 h at the temperature of 20-60 DEG C, then filtration is conducted, and the propolis extract is obtained. And concentrating the filtrate under reduced pressure, and drying to obtain the propolis extract. In a cryptosporidium parvum infected mouse model, the propolis extract shows a remarkable treatment effect; the propolis extract disclosed by the invention can play a role in resisting cryptosporidium parvum through a multi-target and multi-channel mechanism, has multiple biological effects of resisting parasites, inflammation, oxidation, immunoregulation, intestinal barrier protection and the like, and has good development potential and clinical application prospect.

The present invention relates to a combination vaccine comprising the antigens: a) Cryptosporidium parvum gp40; b) inactivated bovine rotavirus; c) inactivated bovine coronavirus; and d) E. coli fimbrial adhesins F5 and F41. In addition, the invention relates to methods for the preparation of the vaccine, methods for vaccinating ruminants against infection by Cryptosporidium parvum, bovine rotavirus, bovine coronavirus and E. coli, and to a kit-of-parts at least comprising a container comprising the at least one antigen according to the invention.

The application discloses application of a micro cryptosporidium cgd6_660 protein as an oocyst outer wall marker protein, and discloses that the inventors find that the cryptosporidium cgd6_660 protein is an oocyst outer wall protein, and the protein is located on the outer surface of the cryptosporidium oocyst wall. The inventors confirm the feasibility of the protein as a detection antigen. The antibody of the protein recombinant protein (as shown in a sequence table SEQ ID N0.2) can be used for immunological detection of unbroken cryptosporidium oocysts (live cryptosporidium oocysts), and the application further provides a specific polypeptide of the protein.

Cryptosporidium parvum is a highly prevalent zoonotic and anthroponotic protozoan parasite that causes a diarrheal syndrome in children and neonatal livestock, culminating in growth retardation and mortalities. Disclosed herein are inhibitors against the enzymatic activity of recombinant CpLDH protein that were identified. The inhibitors were tested for anti-Cryptosporidium effect using in vitro infection assays of HCT-8 cells monolayers. Compounds NSC158011 and NSC10447 were identified to inhibit the proliferation of intracellular C. parvum in vitro, with IC50 values of 14.88 and 72.65 μM, respectively. At doses tolerable in mice, both NSC158011 and NSC10447 significantly reduced the shedding of C. parvum oocysts in infected immunocompromised mice's feces and prevented intestinal villous atrophy as well as mucosal erosion due to C. parvum. These findings have unveiled anti-Cryptosporidium drug candidates that can be explored further for the development of therapeutic agents against C. parvum infections.

The invention relates to an epitope polypeptide of cryptosporidium parvum targeting CD4 + T cells and application of the epitope polypeptide. The amino acid sequence of the epitope polypeptide is shown as SEQ ID NO.1; the epitope polypeptide is a combined long epitope polypeptide of cryptosporidium parvum Gp40 / 15 protein, Gp900 protein and CSL protein; the epitope polypeptide can be applied to a vaccine for preventing or treating cryptosporidiosis parvum, and after a mouse is immunized by the vaccine, not only can CD8 + T cell immune response of the mouse be obviously stimulated, but also stronger CD4 + T cell response is induced to be generated, and the mouse can be induced to generate multifunctional T cells related to an immune protection effect; the levels of IFN-gamma, TNF-alpha and IL-2 generated by multifunctional T cells induced by a long polypeptide immune group have extremely significant differences (P is less than 0.01) compared with various negative controls, which indicates that the long polypeptide epitope vaccine has corresponding biological immunological activity and becomes a novel cryptosporidium parvum infection resistant T cell epitope vaccine or an immunodominance T cell epitope-based mRNA vaccine.

The present invention discloses a direct immunofluorescence reagent, a kit and an application thereof for detecting Cryptosporidium parvum, and belongs to the field of biotechnology. The present invention uses the whole worm protein of Cryptosporidium parvum as an antigen and adopts the hybridoma cell monoclonal antibody technology to prepare a monoclonal antibody against Cryptosporidium parvum; the monoclonal antibody against Cryptosporidium parvum obtained in step S1 is labeled with a fluorescent dye to obtain a direct immunofluorescence detection reagent for detecting Cryptosporidium parvum. The direct immunofluorescence detection method established by using the reagent of the present invention can be used for the detection and identification of Cryptosporidium parvum in clinical stool samples and the environment, and has high sensitivity, strong specificity, good stability, fast diagnosis speed, and good application prospects.

Compositions for preventing or decreasing cryptosporidiosis in animals are disclosed herein. The compositions include sources of hydrogen peroxide and are fed to animals. The animals may be neonatal calves and the cryptosporidiosis may be caused by Cryptosporidium parvum. Methods of preventing or decreasing cryptosporidiosis in animals by feeding the hydrogen peroxide-generating compositions to the animals are also disclosed. Some disclosed methods reduce the number of Cryptosporidium oocysts shed by an infected animal or reduce the infectivity of Cryptosporidium oocysts shed by an infected animal. Some methods include feeding the hydrogen peroxide-generating compositions to animals ultimately afflicted with cryptosporidiosis, and the animals gain weight despite the infection.

The application provides a double-PCR differential detection method for Cryptosporidium parvum and Eimeria bovis, which is used for synchronously differentiating and detecting Cryptosporidium parvum and Eimeria bovis in a to-be-detected sample; the double-PCR differential detection method uses the following primer combination: an upstream primer Cpar-F for Cryptosporidium parvum, a downstream primer Cpar-R for Cryptosporidium parvum, an upstream primer Ebov-F for Eimeria bovis, and a downstream primer Ebov-R for Eimeria bovis. The application also provides application of the double-PCR detection method and a kit for synchronously differentiating Cryptosporidium parvum and Eimeria bovis. The double-PCR differential detection method can simultaneously and specifically amplify two target fragments in the same reaction tube, so that rapid differentiation of two pathogens and mixed infection of the two pathogens can be realized.

The invention discloses a medical application of phellinus igniarius in a medicine for inhibiting the activity of cryptosporidium parvum FBA, provides a medical application of phellinus igniarius in a medicine for inhibiting the activity of cryptosporidium parvum FBA, and discloses that phellinus igniarius can inhibit the activity of rC.pFBA, destroy the normal form of sporozoites, reduce the glycolysis level of sporozoites and relieve the infection of cryptosporidium parvum. The composition can relieve intestinal pathologic symptoms including intestinal villus shortening and erosion, recover the expression quantity of intestinal barrier related proteins and inflammatory factor level, reduce oocyst discharge and the like, and can be prepared into medicines for resisting cryptosporidium parvum.

The invention discloses a traditional Chinese medicine composition capable of relieving diarrhea caused by cryptosporidium parvum as well as a preparation method and application of the traditional Chinese medicine composition, and belongs to the technical field of medicines. The traditional Chinese medicine composition consists of a prescription for clearing intestines and resolving dampness and allicin, the prescription for clearing intestines and resolving dampness is composed of scutellaria baicalensis, coptis chinensis, roasted radix aucklandiae, fried angelica sinensis, sanguisorba officinalis, parched white peony root, angelica dahurica and liquorice. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 10-15 parts of radix scutellariae, 6-10 parts of rhizoma coptidis, 6-10 parts of roasted radix aucklandiae, 10-15 parts of fried radix angelicae sinensis, 10-15 parts of radix sanguisorbae, 15-20 parts of fried radix paeoniae alba, 10-20 parts of radix angelicae, 6-10 parts of liquorice root and 0.02-0.07 part of allicin. The traditional Chinese medicine composition prepared by the invention is efficient, low in toxicity, safe and harmless, and can relieve diarrhea caused by cryptosporidium parvum, relieve intestinal injury, regulate the composition structure of intestinal flora and enhance intestinal barrier and immunity.

The invention discloses a cryptosporidium parvum sporozoites activity detection method and device and application, and belongs to the technical field of biological detection and drug screening. The method comprises the following steps: providing a to-be-detected sample containing dead or live cryptosporidium parvum sporozoites; diluting the to-be-detected sample in a multiple ratio gradient manner and / or performing freeze thawing treatment, and adding the to-be-detected sample into a detection container; an ATP bioluminescence detection reagent is added into the detection container for oscillation treatment, so that cryptosporidium parvum sporozoite is split to release ATP, and a reaction is carried out to generate a luminescence signal; collecting the light-emitting signal, and quantitatively evaluating the survival rate or activity of the cryptosporidium parvum sporozoite according to the intensity of the light-emitting signal. The method provided by the invention is based on the ATP bioluminescence principle, can directly, quickly and quantitatively evaluate the activity of the sporozoite stage in a short time, and can be used for high-throughput screening of large-scale compound drug libraries.

The invention relates to the technical field of biology, in particular to a primer group and a method for detecting cryptosporidium parvum. The primer group for detecting the cryptosporidium parvum comprises primer nucleotide sequences as shown in SEQ ID NO. 1 to SEQ ID NO. 6; according to the present invention, the excrement sample can be directly collected and added into the LAMP reaction reagent integrated in the microfluidic chip, the detection process is automatically completed by the full-automatic detection instrument, the spectrum is analyzed through the spectrum sensor, the collected spectrum signal is comprehensively judged through the AI technology, the full-automatic operation is achieved, the detection time can be controlled within one hour, and the detection accuracy is high. The kit realizes instant diagnosis, does not need professionals and laboratory environments, is portable and light, can be applied to instant detection in scenes such as pet hospitals on site, adopts an innovative LAMP test method, does not need nucleic acid extraction, realizes sample result output, and is low in price.

The invention discloses a lactobacillus johnsonii YS my4 strain with a cryptosporidium infection resisting effect and application of the lactobacillus johnsonii YS my4 strain. The preservation number of the YS my4 strain is CCTCC (China Center For Type Culture Collection) M 20253011. The strain disclosed by the invention has the characteristics of cholate resistance, intestinal juice resistance, strong adhesiveness and capability of inhibiting salmonella and staphylococcus aureus. In addition, the strain has no hemolytic rings, and the strain has no influence on body indexes and can increase the body weight after gavage of suckling rats, so that the strain is proved to be high in safety and can be applied to the fields of medicine and food. According to the invention, the YS my4 strain is intragastrically fed to a pseudo-sterile suckling mouse infected by cryptosporidium parvum, and it is found that the strain can activate immune defense of the intestinal tract of a host by improving the MAPK phosphorylation level of the host in the state of cryptosporidium parvum infection, so that the insect loading amount of the cryptosporidium parvum is remarkably reduced; meanwhile, the YS my4 strain can improve the expression level of tight junction proteins Occludin and CLDN4, relieve pathological damage of ileum tissues and improve the intestinal barrier function, and has important application value in prevention and treatment of cryptosporidium parvum infection.

The invention belongs to the biological field and discloses a cdc42 Gene knockout HCT-8 cell line and its application, cdc42 The taxonomic name of the knockout HCT-8 cell line is Homo sapiens , deposited in Guangdong Provincial Microbial Culture Collection Center, with the deposit number GDMCC NO: 66616; deposit date July 1, 2025; deposit address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, Guangdong Province. The present invention uses CRISPR-Cas9 technology to knock out HCT-8 cells cdc42 Gene to construct cdc42 The gene-knockout HCT-8 cell line was used as a model for Cryptosporidium parvum cell infection.

The application discloses a Cryptosporidium parvum CpDHR1 gene replacement strain and a construction method and application thereof. The application researches and finds that the CpDHR1 gene of Cryptosporidium parvum is related to virulence, the CpDHR1 gene of a high-virulence strain is replaced by the CpDHR1 gene of a low-virulence strain through a gene editing technology, and a Cryptosporidium parvum CpDHR1 gene replacement strain is successfully constructed. Research shows that after the CpDHR1 gene of a high-virulence wild-type Cryptosporidium parvum IIdA20G1 subtype strain is replaced, the strain can still be cultured in vitro, but the in-vitro growth of the strain is significantly reduced, the virulence of the strain is weakened, the oocyst excretion intensity of the strain is weakened, the oocyst load in mice is reduced, the clinical symptoms of the mice are alleviated, the survival time of the mice is prolonged, the virulence of the strain is significantly weakened, the strain can be used for preparing a Cryptosporidium parvum low-virulence vaccine product, and Cryptosporidium parvum infection can be better prevented.

Cryptosporidium parvum is a highly prevalent zoonotic and anthroponotic protozoan parasite that causes a diarrheal syndrome in children and neonatal livestock, culminating in growth retardation and mortalities. Disclosed herein are inhibitors against the enzymatic activity of recombinant CpLDH protein that were identified. The inhibitors were tested for anti-Cryptosporidium effect using in vitro infection assays of HCT-8 cells monolayers. Compounds NSC158011 and NSC10447 were identified to inhibit the proliferation of intracellular C. parvum in vitro, with IC50 values of 14.88 and 72.65 μM, respectively. At doses tolerable in mice, both NSC158011 and NSC10447 significantly reduced the shedding of C. parvum oocysts in infected immunocompromised mice's feces and prevented intestinal villous atrophy as well as mucosal erosion due to C. parvum. These findings have unveiled anti-Cryptosporidium drug candidates that can be explored further for the development of therapeutic agents against C. parvum infections.

The invention relates to an epitope polypeptide of cryptosporidium parvum Gp900 protein and application of the epitope polypeptide, the amino acid sequence of the epitope polypeptide is shown as SEQ ID NO.1, and the nucleotide sequence for coding the epitope polypeptide is shown as SEQ ID NO.2. Experiments prove that the epitope polypeptide has good in-vitro affinity with mouse H-2Db and has certain stability; after the epitope polypeptide is emulsified by an adjuvant to immunize a mouse, not only can CD8 + T cell immune response of the mouse be obviously stimulated, but also polypeptide specific CD4 + T cell response can be induced to be generated; compared with negative control, P is less than 0.01, and the IFN-gamma and the TNF-alpha generated by CD8 + T cells and CD4 + T cells induced by the Gp900 protein have extremely significant difference, so that the Gp900 protein and the related polypeptides thereof have corresponding biological immunology functions, have huge development and application potential in the field of cryptosporidium parvum specific immunotherapy, and have a broad application prospect in the field of cryptosporidium parvum specific immunotherapy. And a new direction is provided for preparing cryptosporidium parvum vaccines based on T cell epitopes.

The invention provides application of a compound in preparation of a medicine for resisting cryptosporidium parvum, and belongs to the technical field of application of chemical raw material medicines. The compound is 5-amino-3-(1-cyano-2-(3-phenyl-4-vinylpyrazole))-1-phenyl-4-cyanopyrazole, the structural formula of the compound is shown as a formula (I): # imgabs0, and it is proved that the compound serves as an inhibitor of cryptosporidium parvum alternating oxidase CpAOX, has a remarkable effect in the aspect of preventing and treating cryptosporidium infection and is worthy of clinical popularization.

The invention discloses a multiple PCR (Polymerase Chain Reaction) primer group and a detection method for simultaneously detecting four kinds of animal-and-human protozoa of goats, which are used for detecting enterosporidium pichia, cystic protozoa, cryptosporidium parvum and giardia duodenalis. The primer group comprises the following primers: a specific primer of enterosporidium pichia, a specific primer of cyst protozoa, a specific primer of cryptosporidium parvum and a specific primer of giardia duodenalis. In addition, the invention also discloses a multiple PCR (Polymerase Chain Reaction) detection method for the enterosporidium pichia, the cystozoon, the cryptosporidium parvum and the giardia duodenalis. Experiments prove that the method has the advantages of rapidness, sensitivity and high specificity, the operation steps of traditional step-by-step PCR are remarkably reduced, the purpose of simultaneously detecting a plurality of pathogen target genes is achieved through one-time reaction, the detection cost is saved while the detection time is shortened, a foundation is laid for accurately and rapidly identifying pathogens, and the method has a relatively great development prospect.

The present invention relates to a compound or a pharmaceutically acceptable salt thereof, a composition containing the compound or the pharmaceutically acceptable salt thereof, a combination comprising the compound or the pharmaceutically acceptable salt thereof and at least one additional therapeutic agent, and the use of the compound or the pharmaceutically acceptable salt thereof, the composition, the combination in therapy, and the use of the compound or the pharmaceutically acceptable salt thereof. For example, the compound is used for treating infectious diseases or treating diseases caused by cryptosporidium.