138 results about "Blood flukes" patented technology

A blood fluke infectious oncomelania detection kit and a detection method thereof belongs to the verminosis transmission medium detecting field. The invention provides a kit and a detection method for detecting blood fluke infectious oncomelania based on a loop-mediated isothermal DNA amplification technology (LAMP). According to the LAMP technology principle, six pairs of specific primers for amplifying a DNA fragment between the 20bp and 231bp of the schistosoma japonicum non-long terminal repeated inverse transcription transposon gene (AF412214) are designed. The LAMP method is constructed for detecting blood fluke gene in the body of the infectious oncomelania and the objective for distinguishing the blood fluke infectious oncomelania from the non-infectious oncomelania is reached. Comparing to the conventional blood fluke infectious oncomelania detection method the method of the invention has advantages of easy and fast operation, sensitive and specific without especial equipment and is suitable for bilharziasis controlling personnel use on site.

The invention discloses gene orders of schistosoma japonicum recombinant multi-epitope antigens BSjGCP-BSj23 and BSjGCP-BSj23-BSj28, a method for expressing and purifying the same, and application thereof in preparing schistosomiasis japonica immunity prevention vaccines and diagnostic reagents. Recombinant multi-epitope nucleic acid vaccines pCMV-BSjGCP-BSj23 and pCMV-BSjGCP-BSj23-BSj28 obtain 14.76 percent and 64.95 percent of worm reduction rates respectively in Kunming mice. The recombinant multi-epitope antigens pGEX-BSjGCP-BSj23 and pGEX-BSjGCP-BSj23-BSj28 obtain 15.7 percent and 57.99 percent of worm reduction rates in immunizing BalB / c mice, and obtain 91.0 percent and 89.9 percent of sensitivities as well as 97.8 percent and 93.4 percent of specificities respectively as diagnostic antigens.

The invention relates to application of a sanguinarine or a chelerythrine for preventing and curing of schistosomiasis. The invention applies the sanguinarine and the chelerythrine on killing an oncomelania, a snail egg, a Japanese blood fluke cercaria, a schistosome and the prevention and cure of schistosomiasis. The inventor discovers that the sanguinarine or the chelerythrine can kill the oncomelania and the snail egg safely and effectively and can kill the Japanese blood fluke cercaria as well as a schistosomulum and an imago of the schistosome under a lower concentration, which has the functions of preventing and curing the schistosomiasis and does not pollute the environment.

The present invention belongs to immunological parasitic disease diagnosis technology. The reagent kit is prepared via dye labeling process and includes labeling antigen solution compounded with 2BLN disperse blue dye, soluble schistosoma japoncium egg antigen, 1% sodium azide and bow serum albumen blocking liquid; and test paper strip with nitrocellulose as chromatographic film detection lines of human sheep antigen IgG and contrast lines of soluble schistosoma japonium egg resisting rabbit antigen IgG on the chromatographic film. The serum to be tested is mixed with the labeling antigen solution and the mixture liquid is tested with the test paper strip.

The invention discloses a fluorescence quantitative PCR reagent box for rapid detection of Schistosoma japonicum, relates to gene detection technology of a kind of pathogen of zoonotic infectious disease, suitable for qualitative and quantitative detection of Schistosoma japonicum. The invention comprises of DNA extracting solution, fluorescence quantitative PCR reaction solution, standard positive template PU-SJ and negative quality control standard sample; the fluorescence quantitative PCR reaction solution contains specific primers and fluorescence probe of Schistosoma japonicum. The invention was accurate in quantitative detection, quick with good specificity and high sensitivity; can identify and detect Schistosoma japonicum and Chinese schistosomiasis; the steps are simple with good repeatability; can qualitatively and quantitatively detect the DNA samples of all three species of Schistosoma japonicum group, can replace traditional aetiology methods, oncomelania crushing method of detecting the snail of schistosome cercariae and larva escape method.

The invention provides a Japanese blood fluke ovum antibody magnetic particle EILSA detecting method, which belongs to verminosis immunodiagnosis technology. The invention adopts the immunodetection theory of antigen sandwich assay method, which uses the surface of magnetic microsphere which is connected with the Japanese blood fluke ovum antigen and has the diameter of 0.1 to 5.0 micrometer as the solid phase reagent; the Japanese blood fluke dissoluble ovum antigen labeled by biological enzyme is used as enzyme reagent. The magnetic separation reagent and the enzyme reagent are respectively combined with the Japanese blood fluke ovum antibody in the sample to form the solid phase- antigen- antibody- enzyme antigen sandwiched immunocomposite. The invention is characterized in that first, magnetic particles are used for replacing traditional enzyme plate ELISA as the solid phase carriers so that the immune reaction is carried on in the condition of liquid phase and the reaction if more abundant and quick; compared with the traditional ELISA method, the method is characterized by the quick detecting speed, the high specificity and the good repetitiveness; second, due to the adoption of bi-antigen sandwich method theory, one kit can be used for the detection of both people and animal Japanese blood flukes.

The invention provides a nano magnetic bead labeled by chicken yolk immune globulin IgY and used for resisting the soluble antigen of schistosoma japonica ovums and a chicken yolk antibody-magnetic bead ELISA (Enzyme-Linked Immuno Sorbent Assay) method for detecting schistosoma japonica circulating antigen. The method comprises the steps of: immunizing a hen with specific antigens of schistosoma japonica with a method of subcutaneous multi-point injection, and extracting, purifying and identifying specific IgY from an egg yolk; and coupling the polyclone IgY on a magnetic bead, and detecting the circulating antigen of the schistosoma by using the magnetic bead-IgY polyclone antibody as a capture antibody and using the specific monoclone IgY antibody as a detecting antibody. The method not only can be used for capturing more antigens by using the magnetic bead and the IgY to increase the sensibility, but also benefits the increase of detecting specificity by using the monoclonal antibody. Thus, the high combination and coordination of sensibility and specificity, required in immunology diagnosis, are realized.

The invention relates to a combined medium and method for testing snail fever. It has the following characters: the medium contains recombination antigen mixture and recombination snail signal protein (rSj14-3-3) monoclonal antibody, and recombination antigen is mixed as recombination snail signal protein (rSj14-3-3) and recombination snail glutathione-s-transferase (rSjGST). The test operation is separately according to indirect enzyme immunosorbent test and double antibody bedded texture method. The invention adopts the two advantage diagnosis antigen molecule and its corresponding monoclonal antigen to form combined testing medium box.

The present invention belongs to the field of biological pesticide, and is especially one kind of high efficiency and low toxicity molluscacide containing ginkgolic acid. The present invention adopts ginkgolic acid as main component, and niclosamide, copper ion and supplementary material to compound composite molluscacide for killing water snail as the unique intermediate host of blood flukes and preventing propagation of schistosomiasis. The test shows that the molluscacide has fast water snail killing effect, no damage to human body and no obvious toxicity on fresh water fish. The molluscacide may be prepared into concentrated liquid preparation or solid preparation for easy production, storing, transportation and use.

The primary objective of the present invention is the development of new mutant forms of the Sm14 protein, for producing a greater production volume. The recombinant proteins here obtained were capable of providing protection against schistosome and fasciola infection. The level of protection of Sm14 recombinant proteins obtained in the present invention was similar to that reached in the parasite saline extract. The mutant proteins of the present invention have reached approximately 100% of renaturation after the heating at 80° C., different from wild forms of the Sm14 protein. Moreover, after storage for 2 months at 4° C., mutant proteins have shown smaller β-structure loss than wild forms that have shown formation with random structure, as demonstrated by the circular dichroism analysis, indicating the success of mutations.

The present invention discloses one new gene coding Japanese blood fluke cell periodic arrest prime, the gene coded polypeptide and the antibody of the polypeptide. The present invention also discloses Japanese blood fluke cell periodic arrest prime of the polypeptide and the application of the polynucleotides as primer or probe, especially the use of the polypeptide and the polynucleotides as diagnosis kit and vaccine.

The invention provides specific antigens of Schistosoma japonicum, which are Schistosoma japonicum immunisin protein Sj50 and calcium bonded protein Sj16, the genes of Schistosoma japonicum immunisin protein Sj50 and calcium bonded protein Sj16 are isolated, these two proteins are expressed through recombination, thus achieving the source of large quantity diagnosis antigens. Sj50 and Sj16 can responsively and specifically detect Schistosoma japonicum antibody, especially Sj50 has the possibility of distinguishing acute and chronic infection, thus the invention has good application prospect.

The invention discloses a Japanese blood fluke protein which has immunogenicity and contains an amino acid sequence shown in SEQ ID NO:1. The invention also discloses an application of the Japanese blood fluke protein in preparation of vaccines for preventing or treating schistosomiasis. The Japanese blood fluke protein disclosed by the invention has a good immune prevention effect on schistosomiasis, and can be prepared into new vaccines for resisting schistosomiasis.

The invention belongs to the immunology field, in particular to a proteantigen molecule-Japanese blood fluke heat shock protein 60KDa (SjHSP60) which is derived from a blood fluke and is capable of increasing CD4 + CD25 + Foxp3 + regulatory T cells and application thereof. The SjHSP60 has a full-length amino acid sequence as shown in SEQ.ID.NO.1, has a series of identical or highly similar cross-reactive T cell epitopes with HSP60 infected by a host. After being used for mouse in vivo immunization or in vitro stimulus to mouse spleen and lymph gland cells, the SjHSP60 can obviously increase CD4 + CD25 + Foxp3 + Tregs. In practical application, the SjHSP60 can effectively relieve inflammatory symptoms and immunopathological effects caused by arthritis, thereby having wide prospects in the aspects of immunological suppression inducement and treatment of immunological diseases.

The invention belongs to the field of photobiology labeling for preventing and treating parasitic diseases, and relates to synthesis of a phosphorescent iridium complex and the purpose of the phosphorescent iridium complex for the fluorescence labeling of schistosome cercaria. The invention aims to realize the synthesis of the phosphorescent complex and perform fluorescence labeling on cercaria. The iridium complex is in the structure that two azotic aromatic compounds as bidentate ligands are matched with an aromatic compound containing dinitrogen in a coordination manner so as to form the iridium complex. The fluorescence labeling method of the cercaria comprises the following steps of: (1) obtaining the fresh cercaria; (2) dissolving the phosphorescent complex with dimethyl sulfoxide so as to accurately compound 1 mmol of a mother solution, and then diluting the mother solution with a PBS buffer solution so as to obtain 5muM of a diluent; (3) performing fluorescence staining at room temperature; and (4) collecting phosphorescence signals for fluorescent imaging. Through design and synthesis, the phosphorescent iridium complex is used as a fluorescent probe to be used for fluorescence labeling of the schistosome cercaria, and the problems of autofluorescence interference and photobleaching in the fluorescent imaging process of the cercaria are successfully solved.

In the invention, a method of hypodermic multi-point injection of specific antigens of blood flukes is used to immunize a hen, the specific IgY is extracted, purified and identified from egg yolks, and an ELISA diagnosis system IgY-ELISA is established by combining the monoclonal antibody technique. The invention establishes a method for detecting circulating antigens of blood flukes, which is novel, highly efficient, sensitive and specific and also has a curative effect evaluation action. Moreover, the invention also initially provides a novel technique and means for detecting other pathogensand diagnosing infectious diseases.

The invention discloses the Schistosoma japonicum antigen gene SjE16 and SjP50, its coding protein and method for recombinating, expressing and separating the protein. The invention also discloses the prevention and cure vaccine of schistosomiasis prepared from antigen SjE16 antigen SjP50 protein. The Schistosoma japonicum antigen SjE16 and antigen SjP50 coding protein and its vaccine have a broad prospect in the prevention and treatment of schistosomiasis.

The invention provides a Schistosoma japonicum gene recombination living vector vaccine which is composed of porcine pseudorabies and a schistosoma japonicum antigen gene; the schistosoma japonicum antigen gene comprises Sj26GST, SjFABP and Sj23 to form monovalent recombination living vector vaccines rPRV / Sj26GST, rPRV / FABP and rPRV / Sj23 as well as multivalent living vector vaccines rPRV / Sj26GST-FABP and rPRV / Sj26GST-FABP-Sj23; the imago reduction rate is between 38.4 and 56.5 percent; the liver egg reduction rate is between 38.8 and 70.6 percent; the vaccine can effectively induce the immunity protection force for resisting the schistosoma japonicum, can generate longer immunity protection effect for one time immunity, does not have pollution or harmful effect to environment and does not cause untoward effect.

The invention relates to schistosoma japonicum micro-ribonucleic acid (miRNA) and application thereof. Particularly, the invention discloses new miRNA which is separated out from schistosoma japonicum, wherein the miRNA can be specifically combined with a corresponding part of messenger RNA (mRNA) to influence the transcription effect of the corresponding mRNA so as to influence the expression of a corresponding gene. The invention also provides application of the identified schistosoma japonicum miRNA, a precursor sequence thereof, antisense oligonucleotide for adjusting the expression of the miRNA and the like in diagnosis, prevention and treatment of the schistosoma japonicum.

The invention relates to the novel use of calcium cyanamide (CaCN2), especially the novel use in preventing schistosomiasis. The disclosed product can effectively eradicate schistosomiasis host oncomelania, eulimidae and amazonian snails. The product can be used to substitute niclosamide.

The invention provides a primer group, probe and kit for detection of Schistosoma mansoni. The primer group includes an upstream primer and a downstream primer, wherein the sequence of the upstream primer is shown in SEQ ID No. 1, the sequence of the downstream primer is shown in SEQ ID No. 2. The primer group for detection of Schistosoma mansoni and a probe and kit for detection of Schistosoma mansoniare has simple and convenient operation. It is verified through tests that when the primer group, probe and kit for detection of Schistosoma mansoni are applied to detection of Schistosoma mansoni, high specificity, high sensitivity, high accuracy and good repeatability are achieved, test results can be obtained within 20 minutes, and the primer group, probe and kit have low requirements fortest conditions and technologies of the test staff and low test cost.

The invention discloses a niclosamide controlled-release dispersing agent for killing schistosoma japonicum cercaria and a preparation method thereof. The niclosamide controlled-release dispersing agent for killing the schistosoma japonicum cercaria is characterized in that the compositions of the niclosamide controlled-release dispersing agent are niclosamide, paradichlorobenzene, floating carriers and a dispersing agent and the main compositions of the niclosamide controlled-release dispersing agent are the niclosamide and the paradichlorobenzene, wherein the weight ratio of the niclosamide to the paradichlorobenzene is between 10 and 30 to between 90 and 70. The niclosamide controlled-release dispersing agent and the preparation method have scientific and reasonable formula and ratio, advanced and simple technique and good effect of killing the schistosoma japonicum cercaria. The controlled-release dispersing agent for killing the schistosoma japonicum cercaria, which is prepared by processing the niclosamide, the paradichlorobenzene, corncob particles and so on as materials, is applied to a pathogen generation water area, releases the niclosamide at a fixed quantity in virtue of sublimation of the paradichlorobenzene, freely diffuses in virtue of the molecular tension and the function of the dispersing agent, reaches the predicted concentration in a water body, has long lasting period, effectively kills the schistosoma japonicum cercaria, and prevents the schistosoma japonicum cercaria from infecting human beings and livestock; and the death rate of the schistosoma japonicum cercaria reaches 100 percent.

Schistosoma japonicum egg antigen (SEA) bone marrow dendritic cells produce an exosome, an exosome is obtained by isolating and purifying and is used to treat a mouse with immune diseases, and then various indexes of this mouse such as body weight change, disease activity index, colon length, tissue lesions and pathological tissue score show that the disease of the experimental mouse is clearly better; it is noted that the exosome produced by japonicum egg antigen (SEA) bone marrow dendritic cells (BMDC) has clear therapeutic effect on immune diseases. The invention belongs to biological therapy, toxic and side effects of existing anti-inflammatory drugs are avoided maximally, the exosome has low toxicity and high efficiency, and a new method for treating immune diseases is provided.

The invention relates to a hybridoma for secreting an anti-recombinant schistosoma japonica enolase specific monoclonal antibody as well as a preparation method and application of the hybridoma, and belongs to the technical fields of gene engineering, preparation of monoclonal antibodies and immunologic diagnosis. The hybridoma JYG-1 capable of secreting the anti-recombinant schistosoma japonica enolase (Sj Enolase protein) specific monoclonal antibody is preserved in the Common Microbiology Center of CCCCM (China Committee for Culture Collection of Microorganisms) and has the preservation number of CGMCC NO. 9910. The hybridoma JYG-1 can secrete the anti-recombinant schistosoma japonica enolase (Sj Enolase) specific monoclonal antibody JYGENmb-1. The monoclonal antibody JYGENmb-1 can be combined with Sj Enolase protein specificity of schistosoma japonica, a sandwich ELISA method for detecting Sj Enolase protein is provided, and the monoclonal antibody can be applied to establishing various immunological methods for detecting Sj Enolase protein in schistosoma japonica infected human and animal blood and excreting secretion.

The invention discloses a serum biomarker for early diagnosis of schistosomiasis japonica. The serum biomarker is phosphatidylcholine and / or palmitoyl choline. The sensitivity and specificity of the marker are both greater than 0.9, and the under-curve area (AUC) is greater than 0.9, which indicates that the two indexes have favorable predictability and can be used for early diagnosis of schistosomiasis japonicum katsurada. The invention further discloses a screening method of the serum biomarker for early diagnosis of schistosomiasis japonica based on metabonomics. A mouse schistosomiasis japonica model is constructed, and the metabonomics analysis is carried out on mouse serum by utilizing an ultra-high performance liquid chromatography-tandem mass spectrometry technology, and characteristic differential metabolites between a normal mouse and a schistosomiasis japonica infected mouse are found and analyzed, namely, the schistosomiasis japonica early diagnosis biomarker is obtained. The screening method has the characteristics of convenience and quickness, can accurately reflect the metabolic spectrum difference between a schistosomiasis japonicum infected mouse and a normal mouse, and is high in specificity.

The invention relates to the field of genetic engineering antibody, and builds schistosoma japonicum immature egg soluble antigen (SIEA) single-chain antibody library for the first time. By screening the SIEA single-chain antibody library through an immuno-affinity screening technique, a specific single-chain antibody aiming at anti-schistosomiasis-japonica effective natural molecular vaccine SIEA26-28kDa is obtained. The obtainment of the specific single-chain antibody provides a foundation for treating schistosomiasis japonica, utilizing the specific single-chain antibody as a probe to screen schistosoma japonicum cDNA libraries and obtaining a corresponding gene sequence aiming at anti-schistosomiasis-japonica effective vaccine molecules more conveniently and rapidly.

The invention discloses a single-chain antibody of an anti-schistosome monoclonal NP11-4 as well as a preparation method and application thereof, and relates to the field of gene engineering and the field of therapeutic drugs against schistosomiasis. The invention is to use a monoclonal antibody NP11-4 located at adult membrane, cercaria memberane, schistosomulum membrane, egg shell and miracidium in egg of Schistosoma japonicum, extract total RNA of hybridoma cells through a gene engineering technology, adopt RT-PCR for proliferation of genes VH and VL, and use overlap-extension PCR to connect the genes VH and VL into a single-chain antibody gene in the form of VH<-linker-VL, which is subsequently connected with a pBAD / gIIIA carrier and cloned into Top10F'of Escherichia Coli to induce a soluble expression of the target gene. The zone from amino acid No.1 to No. 122 of the expressed single-chain antibody of the anti-schistosome monoclonal NP11-4 is a heavy chain gene repertoire of the single-chain antibody (VH), and the zone from the amino acid No.141 to No.254 is a light chain gene repertoire (VL) of the single-chain antibody, wherein the linker between the heavy and light chain gene repertoires consists of 18 amino acids including repeated glycines and serines, and the single-chain antibody has 254 amino acids in full length.

The present invention is niclosamide ethanolamine spray as blood fluke disease preventing and treat medicine for farm cattle and its preparation process. The present invention prepares the niclosamide ethanolamine spray through dissolving niclosamide ethanolamine in 0.5-5 weight portions in cosolvent in 5-10 weight portions, adding penetant in 0.5-2 weight portions and emulsifier in 1-5 weight portions, adding anhydrous alcohol via stirring to 100 weight portions and obtain the niclosamide ethanolamine spray. The niclosamide ethanolamine spray is sprayed to the skin of farm cattle to form one lipophilic protecting medicine layer capable of preventing farm cattle against blood fluke disease in 15-30 days.

The invention provides a water surface floatation sustained release medicament for prevention and cure of schistosomiasis, which is a granular sustained release medicament prepared from medicinal active constituent 5.0-60 wt%, solid state carrier material 30.0-90.0wt%, dispersing agent 0.1-10.0 wt%, coating agent 2.0-10.0 wt%, and hydrophobing agent 0.1-5.0 wt%.

The Japanese blood fluke nucleic acid vaccine consists of eukaryotic expression vector pCD or pBK and blood fluke antigen genes. The blood fluke antigen genes includes Japanese blood fluke antigen genes Sj-23, Sj-26 and Sj-32 and constitute monovalent nucleic acid vaccines pCD-Sj23, pCD-Sj26, pCD-Sj32 and pBK-Sj26 and multivalent nucleic acid vaccine pBK-Sj26-Sj23. The vaccines have blood fluke reducing rate of 35.4-44.4 % and blood fluke egg reducing rate in liver of 39.6-69.0 %, and can induce effectively the immunological protecting capacity to blood fluke. Once immunization has relatively long immunological effect, and the present invention has no environmental pollution, bad influence, and bad reaction.