97 results about "Schistosomiases" patented technology

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 invention discloses a pharmaceutical composition for treating liver diseases, which is prepared by the following active ingredients according to the mixture ratio by weight: liquoric root: white paeony root is equal to 1: 1.1-1: 1.9. In addition, the invention further discloses a preparation method, a use and a quality detection method of the pharmaceutical composition. Clinical observation proves that the pharmaceutical composition can effectively reduce aminotransferase, stop liver area pain and effectively restore liver functions, so that the pharmaceutical composition can be used for treating acute and chronic viral hepatitis and early cirrhosis, and also has better efficacy for patients with advanced schistosomiasis. The medicament with the formula has no toxicity or side effects and can meet the period treatment demand of the patients with the acute and chronic viral hepatitis.

The present invention discloses a medical test paper strip for a fast diagnosis of schistosomiasis japonicum in domestic animals. Said medical test paper strip in accordance with the present invention adopts SEA antibody and SEA coated nitrocellulose membrane as a quality control line and a detection line and is manufactured using a double antigen sandwich method, wherein the SEA antigen is labelled by colloidal gold. The present invention has a remarkable specificity and a high sensitivity, and is operated conveniently, simply and quickly. The present invention has a low manufacturing cost and then is suitable for a mass production.

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 invention provides a kit for detecting the circulating antigen of oriental schistosomiasis, comprising streptavidin-coated magnetic beads, an oriental schistosomiasis resistant soluble egg antigen polyclonal antibody marked by biotin, an oriental schistosomiasis resistant soluble egg antigen specificity monoclonal antibody marked by an electrochemiluminescence reagent, a cleaning solution, a sample diluent, schistosomiasis positive serum, schistosomiasis negative serum and a co-reaction reagent. The invention further provides a method for detecting the circulating antigen of oriental schistosomiasis. In the invention, the kit for detecting the circulating antigen of oriental schistosomiasis is simple, convenient and fast in operation, is suitable for the clinic diagnosis, the curative effect check and the epidemiological investigation at the departments of hospital clinical laboratory, disease prevent and control center, community healthy center and the like, and has very high use value; and the double-antibody sandwich method for detecting the circulating antigen of oriental schistosomiasis has high sensitivity, strong specificity and good repeatability, and is suitable for the detection of batches of specimens and the curative effect check.

The invention discloses siRNA of schistosoma japonicum SjELAV-like 2 genes. The siRNA is selected from one pair of a combination of more than optional two pairs as follows: nucleotide sequences shownas SEQ ID NO.1 and SEQ ID NO.2, nucleotide sequences shown as SEQ ID NO.3 and SEQ ID NO.4, and nucleotide sequences shown as SEQ ID NO.5 and SEQ ID NO.6. The invention further discloses the application of the siRNA of schistosoma japonicum SjELAV-like 2 genes. The siRNA of schistosoma japonicum SjELAV-like 2 genes disclosed by the invention is capable of obviously inhibiting transcription of the schistosoma japonicum SjELAV-like 2 genes and obviously reducing the liver egg hatching rate of schistosomiasis infected mice, and is applicable to preparation of drugs for treating schistosomiasis.

The invention discloses a device for detecting paragonimiasis and preparation method, comprising a label pad, a detecting layer, wherein the labelled detection antigen is on the labelled pad and the detection antigen is the lung fluke ESP protein represented by the amino acid sequence SEQ ID NO.2. The preparation method comprises preparing the lung fluke ESP protein and polyclonal antibody thereof, and preparing the label pad, the detection layer, the sample pad, the absorption pad and the detection device. The detection device prepared by the preparation method has features: high specificity, sensitivity, small cross reaction with the clonorchiasis and schistosomiasis, false positive rate lower than ELISA method, with curative effect evaluating function. The detection result becomes negative in 3-6 monthes after curing the paragonimiasis by effective medicine and the negative rate is higher than the ELISA method and the device for detecting paragonimiasis can accurately, quickly, simply perform diagnosis, differential diagnosis and evaluation of the curative effect with important clinical application value.

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 present invention provides a method of detection of Schistosoma sp. in biological samples through PCR, as well as a method for the diagnosis of Schistosomiasis through the amplification by PCR of the DNA sequence of Schistosoma sp., followed by the separation of the products of the amplification by electrophoresis and the detection by appropriate technique. Moreover, the present invention provides a kit for the diagnosis to be used in the detection of Schistomsomiasis.

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 discloses a urine biomarker for Japanese schistosomiasis early diagnosis. The urine biomarker is one or more of a xanthurenic acid, a naphthalenesulfonic acid, or enanthylcarnitine. Sensitivity and specificity of the marker are greater than 0.9, and an area under the curve (AUC) is greater than 0.9, indicating that the three indicators have better predictability and may be used for the Japanese schistosomiasis early diagnosis. The invention further discloses a metabolomics-based screening method for the urine biomarker for the Japanese schistosomiasis early diagnosis. By constructing a mouse Japanese schistosomiasis model, ultra performance liquid chromatography-tandem mass spectrometry is used to perform metabolomics analysis on mouse urine, and a characteristic differentialmetabolite, that is, the biomarker for the Japanese schistosomiasis early diagnosis, between a normal mouse and a Japanese schistosomiasis-infected mouse is found and analyzed. The screening method is non-invasive, convenient and fast, and can accurately reflect a metabolic spectrum difference between the Japanese schistosomiasis-infected mouse and the normal mouse and have high specificity.

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 relates to an application of a 2-aryl substituted pyrrole compound in a drug for killing bulinus, and belongs to the technical field of schistosomiasis control. The invention particularly provides a chemical structure and preparation method of the 2-aryl substituted pyrrole compound. A molluscicide is prepared with the 2-aryl substituted pyrrole compound as the active ingredient. Themolluscicide has a form of suspension agent, emulsion or microemulsion, and is applied to kill bulinus. When the 2-aryl substituted pyrrole compound is used as the active ingredient of the molluscicide, the application amount for soaking killing is 0.5 mg / L-10.00 mg / L, and the soaking killing time is 24 h-72 h. Experiments prove that the 2-aryl substituted pyrrole compound has the killing activity against bulinus, has the toxicity to aquatic organisms lower than that of conventional snail-killing drug niclosamide, and can be made into the molluscicide applied in the field of schistosomiasis control.

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 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.

The invention relates to the field of biomedicine and discloses a schistosomulum co-culture method by co-culturing schistosomulum and definitive host cells thereof. Proved by experiments, Japanese schistosomulum cultured for 3 days by using the co-culture method of the invention not only has the surface structure, the tegument structure, and the like more similar to a 3-day lung-stage schistosomula growing in a host than a schistosomula cultured for 3 days by using a conventional in-vitro culture method, but also has the gene expression and the soluble proteins characteristics more approaching to the schistosomula growing in the host. The culture method of the invention more approaches to an in-vitro schistosome culture method of in the in-vivo environment of the host, is beneficial to the research on schistosome genomics, post-genomics, transgenic and proteomics and schistosomiasis immunology and lays a foundation for disclosing a growth and development mechanism of the schistosome and the mutual relation between the schistosome and the host.

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 provides genes highly expressed in Schistosoma japonicum schistosomulum and encoded proteins and application thereof. A whole-genome expression profile chip of Schistosoma japonicum is utilized to screen a series of genes highly expressed in Schistosoma japonicum schistosomulum, and antigen proteins encoded by genes which are SjScP27, SjScP80, SjScP84 and SjScP88 can be specifically recognized by serum of patients with schistosomiasis and show obvious positive reactions. ELISA detection proves that the antigen proteins have high sensitivity and specificity for Schistosomajaponicum detection, and can be used for developing diagnostic reagents for schistosomiasis japonica.

The present invention discloses Schistosoma japonicum specific male fluke protein and its gene sequence and specifically combined antibody as well as Schistosoma japonicum preventing and treating vaccine prepared with the specific male fluke protein as immunogen and the application of the specific male fluke protein in preparing medicine for preventing and treating Schistosoma japonicum. Applying the specific male fluke protein can inhibit the sexual maturity of female fluke and disturb the pairing of female and male flukes and the ovipositiom of female fluke, so as to treat schistosomiasis and prevent Schistosoma japonicum diffusion.

The invention relates to a schistosoma japonicum thioredoxin glutathione reductase Sj TGR single-domain antibody and a preparation method thereof, and belongs to the technical field of molecular biology, immunology and pharmacy of a pharmaceutical technology. The invention provides a recombinant phage display library for displaying an Sj TGR protein single-domain antibody, the Sj TGR protein resistant single-domain antibody and a preparation method thereof. The Sj TGR resistant single-domain antibody disclosed by the invention can be specifically combined with an Sj TGR protein and / or has Sj TGR inhibition activity and can be used for preparing a new drug for curing schistosomiasis or a targeting vector of the drug for cure. The schistosoma japonicum thioredoxin glutathione reductase Sj TGR single-domain antibody disclosed by the invention lays a good foundation for developing the new drug for curing the schistosomiasis and a new curing method and has important significance on controlling schistosomiasis prevalence.

The present invention relates to a schistosoma japonicum monoclonal anti-idiotypic antibody NP30 monospecific diabody and a preparation method and applications thereof., which belong to the field of genetic engineering technology and immunotherapy drug preparation. The preparation method utilizes the genetic engineering technology to prepare the schistosoma japonicum monoclonal anti-idiotypic antibody NP30 monospecific diabody and adopts overlap PCR to amplify diabody gene VH-linker-VL (diabody, D for short), the length of linker chooses five amino acid residues, the sequence is GGGGS, and two scFv molecules are forced to mutually form VH-VL pairing, and bound together by a non-covalent bond to form a dipolymer, which is called diabody. The diabody can be used to treat acute schistosome infection and immunoprotection against schistosomiasis.

The invention relates to the technical field of biology and particularly discloses application of a schistosomulum japonicum high-expression gene or coding protein thereof. The invention provides application of schistosoma japonicum katsurada SjScP25 gene, SjScP37 gene or SjScP41 gene, or coding protein thereof or a biomaterial containing the genes in preparation of schistosoma japonicum katsuradavaccine or schistosoma japonicum katsurada medicines, and provides application of the schistosoma japonicum katsurada SjScP37 gene or coding protein thereof or a biomaterial containing the schistosoma japonicum katsurada SjScP37 gene in preparation of schistosoma japonicum katsurada detection reagents or kit or detection of schistosoma japonicum katsurada with nondiagnostic purpose. The schistosomulum japonicum high-expression gene serving as antigen to immunize mice to achieve a relatively good protection effect, can relieve, even resist schistosomiasis infection and can be applied to development of schistosomiasis vaccines. Furthermore, the schistosomulum japonicum high-expression gene or coding protein thereof have high sensitivity and specificity on schistosomiasis detection and can be used in development of schistosomulum japonicum diagnostic reagents.

The present invention discloses a fusion gene of schistosoma japonicum antigen genes, formed DNA vaccine and its preparation method. Said schistosoma japonicum DNA polyvalent vaccine is composed of schistosoma japonicum antigen genes (including sj23, sj FABP, sj26, sj GAPDSH, sj TPI and sj 97) and fusion gene of schistosoma japonicum antigen genes and eukaryotic expression vector with several insertion sites. The fusion gene of 30 schistosoma japonicum antigen genes is obtained by using 6 schistosoma japonicum antigen genes through 'fusion PCR' preparation process. Said fusion gene or antigen gene can be inserted into eukaryotic expression vector so as to obtain the invented schistosoma japonicum DNA polyvalent vaccine.

The invention relates to a schistosome antigen for inducing short-lived antibody response and a schistosomiasis diagnostic kit and a detection method for detecting the antibody response, belonging to the technical field of schistosomiasis prevention and treatment and immunological diagnosis. Protein molecules such as glyceraldehyde-3-phosphate dehydrogenase (SjGAPDH) and the like capable of inducing short-lived antibody response in a schistosomiasis infected person through an immunoblotting technology and a mass spectrometry method are identified. A recombinant SjGAPD protein is prepared by adopting a genetic engineering technology, and an enzyme-linked immunosorbent assay detection kit and detection method for detecting anti-SjGAPDH protein specific short-lived antibody response in the schistosomiasis infected person by the recombinant SjGAPD protein are established to diagnose the schistosomiasis infected person and estimate the treatment effect.