221 results about "Gondii toxoplasma" patented technology

This invention relates to a method for treatment of latent Toxoplasma gondii infection. The invention provides for the use of Hsp90 inhibitors for treatment of latent Toxoplasma gondii infection, particularly in an immunocompromised subject. Also provided is a screening method for identifying compounds useful for treating latent Toxoplasma gondii infection.

The invention discloses a toxoplasma gondii IgM antibody detection kit combined with the FITC-anti-FITC indirect coating technology and the chemiluminescent immunoassay technology, and a preparation method thereof. The kit of the invention is composed of a negative control, a positive control, solid-phase vectors for anti-FITC antibodies, anti-human Mu-chain monoclonal antibodies of FITC markers, toxoplasma gondii antigens which are marked by horse radish peroxidase, chemiluminescent substrates and concentrated washing solutions. The kit of the invention can be used as the aided detection index for prenatal prepotency diagnosis, and has vital significances for improving the birth population quality and doing the family planning and the prepotency well.

The invention discloses a fluorescent quantitative PCR (FQ-PCR) kit for rapidly detecting Toxoplasma gondii, relating to gene detection technology of zoonosis pathogen, and being applicable to qualitative and quantitative detection of Toxoplasma gondii. The invention comprises DNA extracting solution, a standard positive template, a fluorescent quantitative PCR reaction solution, Toxoplasma gondii B1 gene specific primer and a negative quality control standard product. The invention is accurate in quantifying, rapid in detection speed, good in specificity, high in sensitivity, simple in use steps and high in repeatability and can replace traditional etiology detection method.

The invention discloses a screening method of small-molecule inhibitor capable of inhibiting proliferation of toxoplasma gondii. The method utilizes an Alphascreen experiment to primarily select 11 small-molecule inhibitors and one small-molecule accelerant, and the 12 drugs are used for clinical treatment. The 12 drugs are further screened at the cellular level, a plaque experiment proves that GSK J4HCL apparently inhibits the proliferation of toxoplasma gondii, and the value EC50 of the GSK J4HCL to toxoplasmagondii is determined to be about 4.5 muM by virtue of intracellular proliferation experiment. The AlphaScreen competitive inhibition experiment evaluates that the value IC50 of the GSK J4HCL for inhibiting the interaction of TgAtg8 to TgAtg3 is 11.01 muM. The CCK8 experiment is usedfor proving that the value IC50 of the drug for inhibiting the growth of a host cell is 34.359 muM which is far greater than the value EC50 of the drug for the insect, which illustrates that the GSKJ4HCL is a low-toxicity and high-efficiency drug for resisting the toxoplasma gondii.

The present invention provides multiplex PCR primer probes and a kit for detecting pet-derived zoonotic pathogens. Bartonella henselae, toxoplasma gondii and brucella belonging to pet-derived zoonoticpathogens are subjected to gene sequence analysis and comparison, triple fluorescent PCR detection primers and probes suitable for single reaction and simultaneous detection of the three pathogens are designed, and nucleotide sequences of the primers and probes are shown in SEQ ID NO.1-9, respectively. The present invention also discloses a method and a detection kit for detecting the above 3 pathogens. The detection method simplifies detection workload of 2/3 or more of the original, and shortens detection time by about 1 hour, and detection sensitivities of the bartonella henselae, toxoplasma gondii and brucella are 5, 5 and 50 copies, respectively. The multiplex PCR primer probes have no non-specific amplification of 23 common pathogens and canine and cat chromosomes, and show good specificity.

The invention discloses an indirect enzyme-linked immuno sorbent assay (ELISA) kit for detecting canine and feline Toxoplasma gondii antibodies, relates to an immunological detection technology for human and animal infectious diseases and is applicable to qualitative detection of canine and feline Toxoplasma gondii antibodies. The kit comprises a Toxoplasma gondii recombinant antigen GRA7 pre-coated ELISA plate, wherein the sample diluent is 0.01 mol/L of PBS (with the pH of 7.2) containing 0.5 percent of BSA and 0.05 percent of NaN3; the enzyme bonder is a horse radish peroxidase (HRP)-rabbit anti-dog/cat IgG bonder; the substrate developing solution is a TMB developing solution; the stopping solution is a 2mol/L of sulfuric acid solution, positive control and negative control. The kit is high in sensitivity, high in specificity and high in repeatability and can serve as a method for detecting canine and feline Toxoplasma gondii antibodies.

The present invention belongs to the field of biotechnology, and relates to a surface antigen 1 (SAG1) of Toxoplasma gondii human antibody Fab fragment, encoded gene and use thereof. According to the invention, the surface antigen 1 (SAG1) of Toxoplasma gondii human antibody Fab fragment is filtered from a base through establishing a Toxoplasma gondii human immunoglobulin, ELISA, diluting the prothrombin time, sequencing analysis, etc. Through expression purifying and authenticating, the human antigen Fab fragment is authenticated to specifically identify the tachyzoite-bradyzoite recombination SAG1 of Toxoplasma gondii and have higher affinity with the tachyzoite-bradyzoite recombination SAG1 of Toxoplasma gondii, for being identified with the specificity of Toxoplasma gondii tachyzoite-bradyzoite. The human antigen Fab fragment of the invention does not contain Fc segment and does not activate the alexin or cause the histopathological damages of human immune response, etc. when the function of restricting the invasion of Toxoplasma gondii to the host cell is exerted. The surface antigen 1 (SAG1) of Toxoplasma gondii human antibody Fab fragment is safe and reliable when applied for the human body. The antigen medicine for treating toxoplasmosis or the antigen targeted medicine can be prepared.

The invention discloses a dog toxoplasma gondii antibody indirect ELISA detection kit, and belongs to the technical field of biological inspection and quarantine. According to the dog toxoplasma gondii antibody indirect ELISA detection kit, toxoplasma gondii ROP5 recombinant protein is taken as a detection antigen. According to the dog toxoplasma gondii antibody indirect ELISA detection kit, toxoplasma gondii recombinant protein ROP5 is obtained via gene cloning and prokaryotic expression; and a detection method comprises following steps: establishment of an indirect ELISA detection method, specificity testing, sensitivity testing, and comparison with other dog toxoplasma gondii detection kits. Large-scale standardized production of an envelope antigen of the dog toxoplasma gondii antibody indirect ELISA detection kit can be realized; detection specificity is high; sensitivity is high; repeatability is high; result determination is objective; operation process is simple; and large-scale detection can be realized. The dog toxoplasma gondii antibody indirect ELISA detection kit is suitable for dog toxoplasmosis epidemiological investigation, and detection and clinical diagnosis of dog toxoplasma gondii infection; and a rapid, simple, and specific detection method is provided for diagnosis and prevention, and epidemiological investigation of dog toxoplasmosis.

The invention discloses a toxoplasma gondii TgVP1 extracellular region antigen polypeptide, an anti-toxoplasma gondii TgVP1 polyclonal antibody and the application of the polyclonal antibody. The amino acid sequence of the toxoplasma gondii TgVP1 extracellular region antigen polypeptide is as shown in SEQ ID NO: 1 or a sequence as shown in SEQ ID NO: 1 of which the N end is connected with a cysteine. A New Zealand rabbit is immunized by the toxoplasma gondii TgVP1 extracellular region antigen polypeptide as an immunogen so that the anti-toxoplasma gondii TgVP1 polyclonal antibody can be obtained; an identification result indicates that the polyclonal antibody is capable of specifically identifying the toxoplasma gondii TgVP1 protein and can be applied to the detection of the toxoplasma gondii TgVP1 protein in tests such as ELISA, Western blot and immunofluorescence. A powerful tool is provided for the fundamental research of the protein functions and the research of the protein as a potential anti-toxoplasma gondii drug target; in short, the polyclonal antibody is wide in application prospect.

The invention discloses an attenuated live vaccine capable of preventing toxoplasma gondii infection. The vaccine contains a toxoplasma gondii vaccine strain with simultaneous deficiency of the orotidine-5-phosphate decarboxylase gene and the lactic dehydrogenase 1 gene, wherein the nucleotide sequence of the orotidine-5-phosphate decarboxylase gene is as shown in SEQ ID NO:1, and the nucleotide sequence of the lactic dehydrogenase 1 gene is as shown in SEQ ID NO:2. The attenuated live vaccine provided by the invention has the advantages that basically no virulence exists, the immunocompetencefor toxoplasma gondii of animals can be improved, the acute and chronic as well as congenital infection of toxoplasma gondii can be prevented, the vaccine strain cuts off the drug resistance screening label, and thus the vaccine strain has the potential of being prepared into the genetically engineered vaccine for resisting toxoplasma gondii.

The invention discloses a nano antibody for resisting toxoplasma gondii SAG1 as well as a coding gene and application thereof. A VHH chain amino acid sequence of the nano antibody is as shown by SEQ ID NO. 4. The nano antibody comprises two VHH chains. The nano antibody for resisting the toxoplasma gondii SAG1 is obtained by immunizing a camel by virtue of toxoplasma gondii SAG1 antigen, obtainingan anti-SAG1 nano antibody library, and selecting a nano antibody with good performance from the nano antibody library. The nano antibody has high solubility and conformation stability and high antigen affinity and excellent tissue penetration capability, and has high affinity with SAG1 antigen, the affinity constant KD value is 1.66nM, the toxoplasma gondii can be efficiently detected, and the nano antibody can be used for preparing toxoplasma gondii detection kit.

Pyrimidine auxotroph mutants of apicomplexans are provided which are mutated in one of six enzymes of the de novo pyrimidine biosynthesis pathway. Also provided are methods of protecting an animal against infection by apicomplexans by administering a pyrimidine auxotroph mutant and methods for screening for inhibitors of pyrimidine salvage enzymes in apicomplexans.

The invention relates to the field of molecular biology, and provides specific primers for detecting Toxoplasma gondii. The sequence of an upstream primer in the specific primers is as shown in SEQ IDNo. 1; the sequence of a downstream primer in the specific primers is as shown in SEQ ID No. 2. The invention also provides probes for detecting Toxoplasma gondii. The sequences of the probes are asshown in SEQ ID No. 3-6. The invention further provides a kit for detecting Toxoplasma gondii. The kit comprises the primers and the probes for detecting Toxoplasma gondii. The invention further provides a chip for detecting Toxoplasma gondii. The chip comprises the probes for detecting Toxoplasma gondii. According to the invention, gene chip detection is carried out on Toxoplasma gondii by usingthe primers and the probes, and differential diagnosis can be specifically carried out on Toxoplasma gondii infection conditions.

The invention discloses a primer, a probe, a kit and a detection method for detecting pneumocystis carinii and toxoplasma gondii, and particularly relates to a kit and a detection method for double fluorescent quantitative PCR detection of pneumocystis carinii and toxoplasma gondii. According to the invention, the primer pair and the probe are designed by utilizing conserved regions of pneumocystis carinii and toxoplasma gondii, the primer pair and the probe which can specifically amplify pneumocystis carinii and toxoplasma gondii are screened out, the sensitivity is high, the two pairs of primers and the two probes do not interfere with each other, and a sample only containing 1 copy can be accurately detected; by utilizing the primer pair, the probe and the detection method, the contentsof the pneumocystis carinii and the toxoplasma gondii can be absolutely quantified; the kit is simple in structure, safe in detection, rapid and convenient, can be used for detecting the pneumocystiscarinii and the toxoplasma gondii and tracking and monitoring the treatment effect of medicines of a patient, and is beneficial to carrying out epidemiological investigation and preventing and controlling propagation of the pneumocystis carinii and the toxoplasma gondii.

The invention relates to a kit for absolutely quantitatively detecting Toxoplasma gondii based on digital PCR and a detection method thereof; the kit is composed of a microdrop digital PCR kit and a plurality of microdrop generator sheets; the detection method is characterized by comprising: (1) extracting total DNA of a test sample; (2) using the total DNA as a template to carry out PCR amplification; (3) placing the PCR production and microdrop reader to read signals, analyzing experimental data to obtain an absolute content of Toxoplasma gondii in the sample. The kit and method can provide direct quantitation in Toxoplasma gondii detection, have no need for standard curves, have the advantages, such as good operational convenience, high speed and efficiency, good specific sensitivity and low cost, etiological diagnosis on Toxoplasma gondii can be effectively carried out, detection rate is increased, the common problems of existing Toxoplasma gondii detection kits, such as low sensitivity and high misdiagnosis rate, are solved, and the kit and method are suitable for clinical investigations and large-scale disease detection and supervision, related preventive and control measures can be collected in time, and economic loss is decreased.

The invention provides a recombinant bacillus calmette guerin (BCG) vaccine for toxoplamasis and a preparation method thereof. The preparation method for the recombinant bacillus calmette guerin vaccine comprises the following steps of: performing target (TA) cloning on an obtained toxoplasma gondii cyclophilin gene; sequencing and identifying correctly, performing enzyme cutting and recovering target fragments; connecting the target fragments with an Escherichia coli-mycobacterium shuttle expression vector pMV261 and an integrated expression vector pMV361 which are subjected to enzyme cutting reaction respectively; and converting recombinant plasmids to BCG, and screening by resistance and polymerase chain reaction (PCR) to obtain the recombinant bacillus calmette guerin vaccine for the positive toxoplasma gondii. The vaccine is high in heat stability and easy to transport, store and produce, is not needed to be purified and can be directly used for immune protection tests, and a complex process of protein aftertreatment is avoided, so the cost is reduced greatly, and the recombinant bacillus calmette guerin vaccine is suitable for vast rural areas.