159 results about "Schistosoma japonicum" 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 an immunogenic protein in a schistosoma japonicum soluble egg antigen. The immunogenic protein is selected from amino acid sequences shown in SEQ ID NO:2-SEQ ID NO:14. In addition, the invention also discloses a screening method and application of the immunogenic protein in the schistosoma japonicum soluble egg antigen and is mainly characterized by screening the immunogenic protein in the schistosoma japonicum soluble egg antigen through a combined proteomics technology and a serological method, particularly analyzing the immunogenic protein in the schistosoma japonicum soluble egg antigen by utilizing bidirectional electrophoresis, Westernblot (2D-WB) and a mass spectrum method and using the immunogenic protein as a potential target spot for schistosomiasis diagnosis.

The invention provides a schistosoma japonicum katsurada recombinant protein SjSAPLP4 as well as encoding gene and application thereof. The protein has an amino acid sequence shown as SEQ ID NO.2 or an amino acid sequence having a same function, formed by replacing, omitting and / or adding one or more amino acid residues for the amino acid sequence shown as SEQ ID No.2. The invention also provides a gene sequence for encoding the protein. The recombinant protein SjSAPLP4 is good in immunogenicity, can be used as an excellent diagnostic antigen, can be used for preparing a schistosoma japonicum katsurada diagnosis kit having high sensitivity and high specificity, also can be used for preparing an anti-schistosome vaccine and can be used as a potential drug acting target spot to screen the drug for treating the schistosoma japonicum katsurada.

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 discloses a microsatellite locus and an application thereof in schistosome biometrical genetics. The microsatellite locus is provided with a nucleotide sequence selected form SEQ ID NO: 1-17. The microsatellite locus provides a genetic mark which can effectively apply the schistosome biometrical genetics research. The invention also provides a primer of a microsatellite locus sequence by special augmentation. The primer has favorable augmentation effect and unique augmentation product.

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 an automatic collecting and real-time monitoring device for schistosoma cercaria, which is formed by a water carrier, a water suction pump, an enrichment pool, an observation pool and a digital microscopy, and has the working principles that: on the basis of the water surface life habit of the schistosoma cercaria and depending on the drive action of water flow, the schistosoma cercaria is gradually conveyed and enriched into an effective observation range of the digital microscopy in the observation pool, the schistosoma cercaria entering the observation pool can be on-line observed instantly through the digital microscopy, and the states of the schistosoma cercaria can be recorded. The automatic collecting and real-time monitoring device for the schistosoma cercaria can be used for monitoring and surveying schistosoma epidemics.

The invention discloses a small interfering ribonucleic acid (siRNA) of a schistosoma japonicum katsurada membrane-associated progesterone receptor component (PGMRC2) gene. The siRNA is selected from a pair or combination of any more than two pairs of nucleotide sequences shown in SEQ ID NO.1 and SEQ ID NO.2, nucleotide sequences shown in SEQ ID NO.3 and SEQ ID NO.4, nucleotide sequences shown in SEQ ID NO.5 and SEQ ID NO.6, and nucleotide sequences shown in SEQ ID NO.7 and SEQ ID NO.8. The invention also discloses application of the siRNA of the schistosoma japonicum katsurada PGMRC2 gene. By adopting the siRNA of the schistosoma japonicum katsurada PGMRC2 gene disclosed by the invention, transcription of the Sj PGMRC2 gene can be obviously restricted, and 24% of worm reduction rate can be obtained. Therefore, the siRNA of the schistosoma japonicum katsurada PGMRC2 gene can be applicable to preparation of a medicament for treating bilharziasis.

The invention discloses a schistosoma japonicum nucleic acid detection kit based on RPA and a detection method, and belongs to the field of biotechnology. The kit comprises RPA freeze-dried particles, a buffer solution, magnesium acetate, RPA reaction primers and an RPA reaction probe. According to the schistosoma japonicum nucleic acid detection kit based on the RPA and the detection method, the RPA technology is utilized to establish the detection method of schistosoma japonicum, the sensitivity and specificity are equivalent to those of a Q-PCR method and far higher than those of a Kato-Katz method, a miracidium incubation method, an ELISA and an IHA, the whole reaction is completed within 10-20 min, and the result is determined only through changes of a fluorescent curve. The detection kit has the advantages that the sensitivity and specificity are achieved, and the result is easy, convenient and rapid to determine. The detection kit not only can be suitable for bedside diagnosis, but also can be used for field study and environmental assessment.

The invention discloses preparation of a test strip for detection of antibody of bovine schistosoma japonicum katsurada and an application method thereof, which belong to the technical field of diagnosis of animal parasitic diseases. The test strip consists of a sample gasket, an immune probe dry plate, a chromatographic film, water absorbing paper, a PVC (Polyvinyl Chloride) base plate, and an arrow label coating and a handle label coating which are combined. The immune probe dry plate is prepared by forming a covalent bond conjugate by carboxyl of polystyrene latex beads and amido of rabbit anti-bovine IgG antibody protein and absorbing on a polyester film. The chromatographic film is prepared by using a schistosome soluble antigen anchored on a nitrocellulose membrane as a detection line and goat anti-rabbit IgG antibody as a quality control line. The schistosome antibody in bovine serum can be quickly detected. The method is simple and convenient to operate, and the results are intuitional and accurate. The method can be widely applied to diagnosis, general survey and quarantine of bovine schistosome.

The invention discloses an anti-schistosoma japonicum Sj14-3-3 protein monoclonal antibody and application thereof in immune diagnosis, and belongs to the technical field of immune diagnosis of parasitic diseases. The monoclonal antibody against recombinant Sj14-3-3 protein is generated by hybridoma JYQ5C6-1 which has the collection number of CCTCCC201118. The monoclonal antibody against recombinant Sj14-3-3 protein is applied to the detection of an antibody used by schistosoma infected patients or circulating antigen Sj14-3-3 protein in animal blood in schistosoma infected immune diagnosis. The prepared anti-Sj14-3-3 protein molecular specific monoclonal antibody can be used for establishing various immunological diagnosis methods for detecting anti-Sj14-3-3 protein circulating antigens. The time fluorescence resolution method for detecting circulating antigen Sj14-3-3 protein, established by using the anti-Sj14-3-3 protein specific monoclonal antibody, can further improve the accuracy and the positive rate of detecting the schistosoma circulating antigen Sj14-3-3 protein, and has good application prospect.

The invention discloses application of a DNA target sequence of schistosoma japonicum retrotransposon in schistosomiasis diagnosis and particularly provides retrotransposon shown by any one nucleotide sequence selected from SEQ ID NO:1-25 and can be used as a detection marker for schistosomiasis. The invention also provides a pair of primers for specifically amplifying the retrotransposon sequence.

The invention discloses a schistosoma japonicum recombinant antigen. The recombinant antigen is prepared by expressing a recombinant vector of a schistosoma japonicum MRP1 gene segment; the sequence of the MRP1 gene segment is the nucleotide sequence of the amino acid sequence shown in SEQ ID NO.1. The invention further discloses application of the abovementioned schistosoma japonicum recombinantantigen in preparation of products for diagnosing schistosoma japonicum. The schistosoma japonicum recombinant antigen rSjMRP1 is used for diagnosing schistosoma japonicum, is high in sensitivity, high in specificity, and is of important significance on prevention and treatment of schistosomiasis.

The invention discloses primers, probe, kit and method for the RPA-LFD visualization rapid detection of Schistosoma nucleic acid. Sequences of the primers are as shown in SEQ ID NO.1-2, and a sequenceof the probe is as shown in SEQ ID NO.3. The RPA primers and probe are designed with the SjCHGCS19 gene as a target sequence, and the sensitive and rapid visualization detection of the schistosome nucleic acid is realized by using a RPA amplification technology combined with lateral flow chromatography test strip method. The detection method of the invention has a detection limit of 1 fg for Schistosoma japonicum genome DNA, and is expected to be used in the general detection of Schistosoma mansoni and Schistosoma haematobium. The method can detect circulating nucleic acid of Schistosoma in the serum of mice at an early stage of infection, the operation is simple and fast, no special equipment is required, a reaction temperature is close to room temperature, results can be observed with the naked eye, and the realization of the early and sensitive and rapid detection of intermediate hosts of Schistosoma on site, and the timely and sensitive monitoring of environments with high schistosomiasis transmission risks are facilitated.

The invention discloses siRNA of a schistosoma japonicum katsurada SjFrzb2 gene. The siRNA has nucleotide sequences as shown in SEQ ID NO. 1 and SEQ ID NO. 2. The invention also discloses an application of the siRNA of the schistosoma japonicum SjFrzb2 gene. The siRNA of a schistosoma japonicum SjFrzb2 gene can obviously inhibit the transcription of the SjFrzb2 gene, the insect body number of schistosomiasis infected mice, the number of liver eggs, the liver egg contribution rate and the egg hatching rate of each female insect can be remarkably reduced, a certain pathological influence is alsocaused to the integument structure of the insect and the cell morphology of the gonad, and the product is suitable for being prepared into drugs for preventing and treating schistosomiasis.

The invention provides an anti-schistosoma japonicum Sjp40 chicken egg-yolk antibody which can be specifically bound with schistosoma japonicum Sjp40. The anti-schistosoma japonicum Sjp40 chicken egg-yolk antibody is capable of identifying the schistosoma japonicum Sjp40 through antigen specific binding activity detection thereof; the OD value of the anti-schistosoma japonicum Sjp40 chicken egg-yolk antibody is decreased as the antibody dilution is reduced; the anti-schistosoma japonicum Sjp40 chicken egg-yolk antibody has dependence on antigen concentration; the antibody is obtained by a method comprising the following steps of: (1) collecting eggs of healthy SPF (Specific pathogen Free) grade laying hen which is primarily immunized with purified Sjp40 antigen previously, and primarily extracting IgY from the eggs by the method of diluting ammonium sulfate precipitate with water; (2) further purifying the IgY orderly through a dialysis bag and a DEAE-FF (Diethylaminoethyl-Fast Flow) anion exchange column; (3) performing analysis and identification on the IgY through polyacrylamide gel electrophoresis, Western blotting and ELISA (Enzyme Linked Immunosorbent Assay), wherein the antibody is a chicken egg-yolk immune globulin (IgY) special for the schistosoma japonicum Sjp40; and therefore, a new idea is provided for early diagnosis and prevention and treatment of the schistosoma japonicum.

The invention relates to a medicament used for killing oncomelania and schistosoma japonicum cercaria, in particular to an environmental-protection typed niclosamide slow releasing block and a preparation method thereof. The best proportioning of the slow releasing block of the invention according to the weight portions is 50 portions of niclosamide ethanolamine salt wettable powder, 30 portions of sawdust, 20 portions of perlite and 3 portions of polyvinyl alcohol for the upper floating type and is 50 portions of niclosamide ethanolamine salt wettable powder, 20 portions of sawdust, 30 portions of red soil and 3 portions of polyvinyl alcohol. The invention uses the niclosamide recommended by WHO as the main medicament; the adopted carrier material is sawdust which has low price and causes no environmental pollution; more than 6,000m<2> can be released in the operation of two persons each day, thus saving the niclosamide by 50% and the cost by 40%.

The invention discloses schistosoma japonicum katsurada recombinant antigen. The recombinant antigen comprises an amino acid sequence of schistosoma japonicum katsurada PGMRC2 protein indicated by SEQ ID No.1. The invention also discloses a preparation method of schistosoma japonicum katsurada antigen and application of the antigen in preparing a vaccine or a medicament for preventing or treating the schistosoma japonicum katsurada disease. By adopting the schistosoma japonicum katsurada recombinant antigen, the specificity IgG, IgG1 and IgG1a antibodies for resisting the recombinant antigen can be induced in a mice body, the worm reduction rates of 22.08 percent and 20.097 percent are respectively induced in two animal protection experiments, so that the recombinant antigen is suitable for being used as a candidate vaccine for resisting the schistosoma japonicum katsurada and has good application prospect.

The invention belongs to the field of biotechnology and particularly relates to a schistosoma japonica vaccine salmonella secretion expression vector, which is a recombinant salmonella expression vector that is controlled by a bacterial promoter, guided by effector protein secreted by salmonella and secretion signal of the effector protein and used for secretion expression of schistosoma japonica antigen. The schistosoma japonica vaccine salmonella secretion expression vector can induce secretion expression of a schistosoma japonica vaccine in a hypoxic microenvironment in antigen presenting cells, can exist in vivo stably and continuously and is insusceptible to being lost. The schistosoma japonica vaccine salmonella secretion expression vector, by using an attenuated strain as a carrier, can perform the antigen carrying of the recombinant vaccine representing bacteria by means of oral taking. The schistosoma japonica vaccine salmonella secretion expression vector can be used for preparing schistosoma japonica vaccine for oral taking for preventing and treating schistosomiasis.

The invention discloses a preparation method of a Schistosoma japonicum calcium-binding EF-hand domain containing protein (SjEFCAB) recombinant antigen protein. The preparation method comprises the following steps of: designing SEQ ID NO.3 or a complementary chain 5' thereof as a primer and SEQ ID NO.4 or a complementary chain 3' thereof as a primer, and carrying out amplification on a Schistosoma japonicum SjEFCAB gene sequence; establishing and identifying Schistosoma japonicum SjEFCAB recombinant plasmids; and carrying out induction expression, purification and the like on recombinant protein. Besides, the invention further discloses an application of the Schistosoma japonicum SjEFCAB recombinant antigen protein prepared by using the method in preparation of products for detecting a serum antibody of a Schistosoma japonicum patient. Enzyme linked immunosorbent assay (ELISA) proves that the Schistosoma japonicum SjEFCAB recombinant antigen protein has higher sensitivity and specificity when used for diagnosing the Schistosoma japonicum, is a potential candidate target of diagnosis, and can be used as a target antigen for diagnosing the Schistosoma japonicum.

The invention discloses a schistosoma japonicum recombinant antigen, and the recombinant antigen comprises an amino acid sequence, shown as SEQ ID NO. 1, of schistosoma japonicum phosphoglyceric kinase. The invention also discloses a preparation method of the schistosoma japonicum recombinant antigen and application of the schistosoma japonicum recombinant antigen to prepare vaccines or medicines for preventing or controlling schistosomiasis. In mouse immunization experiments, the schistosoma japonicum recombinant antigen is capable of inducing a mouse to generate specific IgG, IgG1 and IgG2a antibodies capable of resisting the recombinant antigen in vivo at a relatively high level, and in animal protection experiments, through induction of the schistosoma japonicum recombinant antigen, the insect reducing rate is 34.5% and the ovum reducing rate is 32.2%. The experiment results prove that the recombinant antigen is applicable as a vaccine candidate for resisting schistosomiasis and has extremely good application prospect.

The invention discloses a LF-RPA method for quickly identifying schistosoma japonicum katsurada, schistosoma mansoni and orientobilharziasis and application thereof and further discloses a quick detection kit and a detection method. A primer pair, probe and quick detection kit for detecting the schistosoma japonicum katsurada, schistosoma mansoni and orientobilharziasis are established on the basis of molecular biology. The primer and probe can be used for specifically amplifying schistosoma japonicum katsurada, schistosoma mansoni and orientobilharziasis, the kit for quickly detecting nucleicacids of schistosoma japonicum katsurada, schistosoma mansoni and orientobilharziasis has the sensitivity higher than that of PCR, has strong specificity and high detection speed, and is extremely simple and convenient in result determination operation. The detection method and detection kit combined with an existing nucleic acid quick extraction method can achieve the aim of quickly identifyingand detecting schistosoma japonicum katsurada, schistosoma mansoni and orientobilharziasis.

The invention belongs to the field of biogen. About 510000 people suffer from Schistosoma japonicum in China, and 65 thousands of people are threatened. The development of an effective vaccine for resisting the Schistosoma japonicum is an important strategy. The invention aims to provide a candidate vaccine molecule for resisting Schistosoma japonicum and an application thereof in preparing recombinant vaccine for resisting Schistosoma japonicum. The invention provides a Schistosoma japonicum zinc finger protein coding gene SjZFP1 as well as a nucleotide sequence thereof and a corresponding coding amino acid sequence thereof disclosed in SEQ IDNO:1. The invention also provides an application of the SjZFP1 gene in preparing the recombinant vaccine for resisting Schistosoma japonicum; the candidate antigens of the recombinant vaccine for resisting the Schistosoma japonicum prepared by utilizing the gene uses an antigen displayed by T7 bacteriophage and a recombined SjZFP1 gene antigen which both have higher immunoprophylaxis effect.

The invention discloses a polypeptide for inhibiting joint inflammations in mice and application thereof. The micro-molecule polypeptide SJMHE1 originated from the egg and imago antigens of Schistosoma japonicum consists of 24 amino acids, is a T cell epitope shared by human and mouse HSP60 molecules, and can prevent and treat collagen-induced arthritis in mice. The polypeptide of the invention has the beneficial benefits that the polypeptide can be used for treating rheumatoid arthritis and has potential new drug development value.

The invention belongs to the technical field of genetic engineering of enzymes and in particular relates to a mutant of schistosoma japonicum glutathione-S-transferase and an application thereof. The invention provides a mutant enzyme schistosoma japonicum glutathione-S-transferase (sj26GST) for changing the schistosoma japonicum immunizing protection. Compared with a wild type, the single mutant enzyme obtained by mutating the glutathione-S-transferase from 26kDa of schistosoma japonicum (Schistosoma japonicum) has the advantages that the protectiveness of the mutant enzyme of sj26GST is improved, the IgG1 level of the mutant enzyme is obviously higher than that of a recombinant wild type sj26GST protein, the worm reduction rate is improved from the original 34 percent to 40 percent, and the egg reduction rate is improved from 37 percent to 40 percent. The recombinant antigen has good immunogenicity and is suitable to serve as an anti-schistosomiasis candidate vaccine.

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 discloses a schistosoma japonicum katsurada recombinant protein. The schistosoma japonicum katsurada recombinant protein is prepared through expression of a schistosoma japonicum katsurada Vamp2 gene-containing recombinant vector. The invention also discloses a use of the schistosoma japonicum katsurada recombinant protein as a schistosoma japonicum katsurada diagnosis antigen and a use of the schistosoma japonicum katsurada recombinant protein in preparation of a vaccine or drug for preventing or treating bilharziasis. In a mouse immunization experiment, the schistosoma japonicum katsurada recombinant protein can induce generation of anti-recombinant protein specific IgG, IgG1 and IgG2a antibodies in a mouse and an antibody generation level is high. An animal protection experiment result shows that the recombinant protein has a potential as an anti-bilharziasis candidate vaccine and a novel drug target. A diagnosis antigen effect assessment experiment result shows that the recombinant protein has a potential as a diagnosis antigen and has a wide application prospect.

The invention discloses siRNA of a schistosoma japonicum SjLwr gene. The siRNA has nucleotide sequences as shown in SEQ ID NO.1 and SEQ ID NO.2; or nucleotide sequences shown in SEQ ID NO.3 and SEQ IDNO.4, or nucleotide sequences shown in SEQ ID NO. 5 and SEQ ID NO. 6. The invention also discloses an application of the siRNA of the schistosoma japonicum SjLwr gene. The siRNA of the schistosoma japonicum katsurada SjLwr gene can obviously inhibit transcription of SjLwr gene, can obviously reduce schistosome infected mouse insect body number, liver egg number, and liver egg contribution rate and egg hatching rate of each female insect, has certain pathological influence on insect integument structure and gonad cell morphology, and is suitable for preparation of drugs for prevention and treatment of schistosomiasis.

The invention provides a schistosoma japonicum katsurada recombinant protein SjSAPLP5 as well as an encoding gene and an application thereof. The protein provided by the invention has amino acid sequence as shown in SEQ ID No.2 or has amino acid sequence which is formed by replacement, deletion and / or addition of one or more of amino acid residues and has the same function. The invention also provides the encoding gene for encoding the protein. The recombinant protein SjSAPLP5 provided by the invention is excellent in immunogenicity, can be served as an excellent diagnosis antigen, is used for preparing a schistosoma japonicum katsurada diagnostic kit with high sensitivity and high specificity, and is also used for preparing an anti-schistosome vaccine and the drug served as a potential drug effecting target spot and is used for screening and treating the schistosoma japonicum katsurada diseases.