81 results about "Schistosoma" patented technology

The invention relates to the technical field of biology, in particular to an antigen chip used for detecting human parasites antigens in serum and a preparation method thereof. The invention spots antigen array of parasites which are seriously harmful to human body, Schistosoma, Lung fluke, Clonorchis sinensis, Cysticercosis, Sparganum mansoni, Trichina, Angiostrongylus cantonensis, Toxoplasma gondii, and the like, on a solid phase membrane carrier, and specific gamma immunoglobulin (IgG) of human parasites in serum is detected by a specially made vertical flow chip detect device, and the detecting marker is colloid gold second antibody conjugate or colloid gold protein A conjugate of fresh color. IgG antibodies in serum specially bind with the antigens on the membrane in percolation process layer upon layer, and human IgG which specially binds with the antigens is detected by the colloid gold marker. Detecting of a plurality of parasites antigen indexes in serum sample can be finished within several minutes. The device and the method have the advantages of high throughput, simple operation, convenient use and being suitable for parasitic diseases clinical test and serum-epidemiological investigation.

The invention provides a kit for detecting circulating antigen indirect hemagglutination of schistosomiasis and a manufacturing method thereof. The kit is composed of freeze-drying anti-SEA-IgY sensitized erythrocyte, sensitized erythrocyte diluent, sample diluents, positive comparison products and negative comparison products. The method for manufacturing the kit for detecting circulating antigen indirect hemagglutination of schistosomiasis comprises the steps of preparing a schistosoma japonica soluble antigen, performing antigen immunization and collecting eggs, extracting and purifying specific anti-SEA-IgY, preparing hydroformylated red cells, preparing tanned red cells, preparing a specific anti-SEA-IgY antibody sensitized erythrocyte, and preparing freeze-drying red cells. The kit in the invention has the advantages of simple and convenient operation, rapidness, high sensibility, strong specificity and good repeatability.

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 the siRNA of SjLGL gene of schistosoma japonica. The siRNA may be one pair or any more than two pairs of nucleotide sequences represented by SEQ ID No.1 and SEQ ID No.2, nucleotide sequences represented by SEQ ID No.3 and SEQ ID No.4, and nucleotide sequences represented by SEQ ID No.5 and SEQ ID No.6. The invention also discloses the use of the siRNA of the SjLGL gene of schistosoma japonica. The siRNA of the SjLGL gene of schistosoma japonica can obviously inhibit the transcription of the SjLGL gene, obviously lower the hatchability of the eggs of insects in livers of mice infected with schistosomes and is suitable for preparing medicines for treating schistosomiasis.

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 discloses high throughput screening of important schistosoma japonicum antigens and application of the antigens in schistosomiasis diagnosis. Specifically, the invention provides important schistosoma antigens and application thereof in schistosomiasis diagnosis and vaccine preparation. Based on a high throughput functional protein screening technology of glutathione S transferase (GST) fusion expression, 24 positive antigens capable of being identified by the serum of a schistosomiasis patient are acquired, wherein 8 antigens represent strong positive reaction. Schistosomiasis serologic detection shows that the positive antigens for detecting schistosoma have high sensitivity and specificity. The invention also provides a corresponding method and a kit for diagnosing and monitoring the schistosomiasis.

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 an oncomelania-and-schistosoma-preventing water bank zone revetment structure. The oncomelania-and-schistosoma-preventing water bank zone revetment structure is characterized in that a herbaceous plant isolation area is arranged below the high water level of a revetment, a soil-fixing erosion-preventing dike dam area is arranged above the high water level of the revetment, and a woody plant oncomelania-expelling area is arranged on one side, far away from water, of the outside of the soil-fixing erosion-preventing dike dam area. Oncomelania-preventing water-tolerant herbaceous plants are planted below the normal water level of the herbaceous plant isolation area, oncomelania-preventing hygrocolous herbaceous plants are planted at the position above the normal water level and below the high water level, and the wide close-planting mode is adopted by all the plants. The soil-fixing erosion-preventing dike dam area is laid with ecological revetment bricks in a building mode, and the width of the soil-fixing erosion-preventing dike dam area is 2-3m. The woody plant oncomelania-expelling area is of a forest belt structure which is composed of hygrocolous trees and hygrocolous shrubs in a mixed mode, and the purpose of tall-and-short matching, the purpose of space combination, the purpose of complementation between the advantages and the disadvantages, and the purpose of oncomelania-expelling symbiosis can be achieved. According to the oncomelania-and-schistosoma-preventing water bank zone revetment structure, perfect combination between water bank zone water conservancy engineering and forestry schistosoma prevention is achieved through thorough engineering design and scientific plant arrangement, and the oncomelania-and-schistosoma-preventing water bank zone revetment structure can play a remarkable role in expelling oncomelania and preventing schistosoma, and can have the advantages of being ecological and attractive, improving the effect, and being multifunctional.

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 provides a carbazole alkamine compound and a preparation method thereof and application for a parasitic disease resistance aspect. Specifically, the invention provides a racemic modification of the compound shown as type (A) and an enantiomer or salt capable of being accepted by pharmacy of the enantiomer. The compound has excellent anti-schistosoma and anti-hydatid cyst activity. According to the carbazole alkamine compound and the preparation method thereof and the application for the parasitic disease resistance aspect, the design is reasonable, the source of used raw materials is wide, the preparation method is easy and convenient and suitable for utility and can be applied to parasitic disease resistance drug preparation. Type A (please see the specifications for the chemical structural formula)

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 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 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 provides a method for controlling propagation of schistosomiasis in lake regions. The method is characterized in that one or more or all of capsella bursa-pastoris, crescent euphorbia, wild carrot, sedge, Indian mock-strawberry herb, dandelion, clover, tokyo violet herb, artemisia selengensis turcz, field thistle, crescent euphorbia, wormwood, buttercup, wild carrot and the like aregrown on the lake regions. The method uses allelopathy of herbaceous plant in lake regions to control schistosoma miracidium to infect oncomelania so as to achieve the aim of controlling the schistosomiasis in lake regions. The invention discloses that certain plants collected from breeding place of the oncomelania, such as the capsella bursa-pastoris, the crescent euphorbia, the wild carrot, thesedge, the Indian mock-strawberry herb, the dandelion, the clover, the tokyo violet herb, the artemisia selengensis turcz, the field thistle, and the like, have obvious rejection on Japanese schistosoma miracidium; and soak solutions of other plants, such as the crescent euphorbia, the wormwood, the buttercup, the wild carrot, and the like, have obvious killing effect on miracidium. The method has the advantages that the aim of protecting human from infection of schistosomiasis can be realized without killing the oncomelania, and has low cost and no environmental pollution.

The invention provides a method for detecting DNA of schistosoma through a fluorescent probe. An amplification system and the DNA of the schistosoma are mixed, isothermal amplification is carried out to obtain an amplified product, and the amplified product is subjected to real-time detection through a fluorescent probe method. The amplification system comprises the components of a Tris buffer solution, potassium acetate, magnesium acetate, dithiothreitol, polyethylene glycol, ATP, dNTPs, phosphocreatine single-strand binding protein, recombinase, UvsY protein, exonuclease, the fluorescent probe, DNA polymerase and a schistosoma DNA specific primer. The temperature of isothermal amplification is 35-45 DEG C. The time of isothermal amplification is 5-20 min. The method does not rely on a thermal cycler, operation is easy, the DNA of the schistosoma can be detected at the low temperature (35-45 DEG C), the time used for amplification is short (5-20 min), and the method is suitable for primary and on-site detection.

The invention provides application of near infrared xanthene fluorescent dye in fluorescent labeling of adult schistosoma. The near infrared xanthene fluorescent dye has the structure shown in formulaI. In the application, the near infrared xanthene fluorescent dye (NIR-RB) with the structure of the formula I has an emission peak located in a near infrared region and has good light transmittancein living body fluorescence imaging. The NIR-RB can perform fluorescent labeling imaging on the adult schistosomiasis; meanwhile, the fluorescence intensity of the adult schistosoma is quantitativelyanalyzed, the signal-to-noise ratio (S / N) of a fluorescent signal in a worm body region and a background region is 10, which indicates that the NIR-RB used for the fluorescence imaging of the adult schistosoma has a high signal-to-noise ratio.

The invention belongs to the field of biological optical labeling of verminosis, mainly synthesis of a near-infrared fluorescent dye and application of the near-infrared fluorescent dye in adult schistosoma fluorescence labeling. The near-infrared fluorescent dye has the conjugated diene bond structure, and more particularly, the molecular structure is connected through three olefinic bonds; the hydrophilic groups are sulfo (-SO3H) and carboxyl (-COOH); and the near-infrared fluorescent dye can be used for adult schistosoma fluorescence labeling imaging. The invention implements design, synthesis and fluorescence labeling on adult schistosoma by using the near-infrared fluorescent dye. By using the conjugated system, the emitted light of the near-infrared fluorescent dye is in the near-infrared region; and the hydrophilic sulfo and carboxyl are combined for water solubility modification. The near-infrared fluorescent dye is used for adult schistosoma fluorescence labeling, and effectively solves the problems of poor fluorescent dyeing effect, background fluorescence interference and poor light penetrability in the adult schistosoma fluorescence imaging.

The invention relates to the field of parasitic disease media detection, in particular to a schistosoma japonicus detection method and a kit and a primer thereof. The kit comprises an LAMP primer set, a reaction mixture and a DNA polymerase. The reaction mixture contains hydroxynaphthol blue as an indicator. The LAMP primer set comprises an outer primer pair formed by F3 and B3 and an inner primer pair formed by FIP and BIP; the LAMP primer set further comprises an LF loop primer. The kit utilizing the detection method can detect the schistosoma japonicas efficiently, quickly and specifically.

The invention discloses a schistosoma japonicum RPA molecular detection method. The method comprises the following steps: 1) designing a pair of specific primers and a probe according to a sequence ofa schistosoma japonicum SjCHGCS19 gene, and a nucleotide sequence of the schistosoma japonicum SjCHGCS19 gene is shown in SEQ ID NO: 1; 2) extracting the total DNA in a sample to-be-tested, and performing an isothermal amplification reaction of the recombinase polymerase; and 3) detecting an amplified product. The method has high specificity and sensitivity for the detection of schistosoma japonicum, and can detect schistosoma japonicum by sampling feces or whole blood in a short time after the infection of the animal, and facilitates early diagnosis and early warning of schistosoma japonicumas well as early laboratory identification and screening.

The invention provides application of let-7b and rSjp40 (recombinant Schistosoma japonicum p40) in preparation of a drug for preventing or treating schistosoma infected hepatic fibrosis. The application is mainly shown as follows: (1) rSjp40 up-regulates expression of let-7b and promoter thereof in a human LX-2 cell line; (2) let-7b participates in the activation process of inhibiting human hepatic stellate cells (HSCs) by the rSjp40; and (3) the rSjp40 can promote increase of expression of let-7b in hepatic tissue and is in negative correlation with expression of collagen. By the function andmolecular mechanisms of rSjp40 in inhibiting the let-7b in the activation process of the HSCs, schistosoma infected hepatic fibrosis is inhibited at the gene level, the application of the let-7b andrSjp40 in preparation of the drug for preventing or treating schistosoma infected hepatic fibrosis is obtained, and the problem of schistosoma infected hepatic fibrosis is expected to be solved in thefuture.

The invention discloses a specificity detection primer, a detection kit and a detection method for schistosoma japonicam sex, an upstream primer Tsexu2 nucleotide sequence of the specificity primer is 5'-ACGTTAGATACTGCTGTTCA-3, a downstream primer Tsexd2 nucleotide sequence is 5-ATATTGTTCCAAGTACGCAT-3'. In the invention, PCR amplification is carried out on the mould DNA to be detected by the specificity detection primer, the agarose gel electrophoresis is carried out on the amplifying product which is then observed under the ultraviolet light, the amplification strip appears in the positive result, which does not appear in the negative result. In the invention, the specificity detection primer is designed according to female worm specific sequence data of the schistosoma japonicam to establish a rapid, specific, sensitive PCR method used for schistosoma japonicam epidemiology and sex identification. The kit of the invention has simple procedures, strong specificity of the method, high sensitivity and objective result determination, and can be used for sex specific diagnosis of schistosoma japonicam of human and animals.

The invention provides a specific-antigen preparing method used for detecting schistosoma japonicum antibodies by an indirect hemagglutination (IHA) method. The method includes the following steps: step one, preparing schistosoma-specific antigens, wherein in a step 1, soluble adult antigens of schistosoma japonicum are prepared, in a step 2, soluble cercaria antigens of the schistosoma japonicumare prepared, in a step 3, soluble egg antigens of the schistosoma japonicum are prepared, in a step 4, the antigens are treated by sodium periodate, and in a step 5, the soluble adult antigens of theschistosoma japonicum, the soluble cercariae antigens of the schistosoma japonicum and the soluble egg antigens of the schistosoma japonicum after treating of the sodium periodate are proportionallymixed according to a volume ratio of 1:0.5:2, that is, specific antigens are obtained; and step two, preparing an indirect hemagglutination reagent by the specific antigens. According to the method, the crudely prepared soluble antigens of schistosoma japonicum adults, cercariae and eggs are proportioned and mixed after treating of the sodium periodate, the diagnostic reagent is prepared, and thereagent reduces cross-reaction with other diseases, and maintains original high sensitivity and specificity at the same time.

The invention discloses a method for preparing a positive oncomelania of schistosoma. The method comprises the following steps: co-culturing myracidium incubated by schistosoma eggs and sf9 cells to obtain mother sporocyst developed by the myracidium; injecting the mother sporocyst into a negative oncomelania body under microscopy; and continuously feeding the oncomelania containing injected mother sporocyst, and detecting whether the oncomelania is positive or not to obtain the positive oncomelania of the schistosoma. By the method for preparing the positive oncomelania of the schistosoma, cercaria released by the cultured positive oncomelania has the same infectivity and the same biological characteristics with the schistosoma. The method provides an efficient and convenient in vitro manual operation method for studying the genetic functions of the schistosoma.

The invention discloses application of an IRF4 gene in resisting schistosoma infection, in particular to application of the IRF4 gene in preparing or screening a medicament for resisting the schistosoma infection. Meanwhile, the invention also discloses a medicament for resisting the schistosoma infection. The medicament comprises an agent capable of inhibiting or silencing the expression of the IRF4 gene. The invention utilizes IRF4 gene knock-out mice and wild mice to carry out experiments, finds that the IRF4 gene deletion can effectively inhibit the formation of liver granuloma of the micewith schistosoma infection, and obviously improves the liver function of the mice with schistosoma infection. The invention provides a new treatment target for resisting the schistosoma infection, provides a new thought for preparing and screening the medicament for resisting the schistosoma infection, and has great medicinal prospect.

Schistosomiasis mansoni is caused by flukes called Schistosoma(es) that enters the human body through the skin in Schistosoma infested waters. The Schistosomes travel from the skin into human blood vessels where they mate, produce antigen containing eggs that travel from the blood vessels into the small intestines, where they are released in the human feces. Male and female Schistosome mates in human blood vessels, male Schistosomes secrete a protein called TGR β protein to the Trk receptor sites on the females Schistosomes membranes. The process stimulates the formation of chemical SmInAct in female Schistosomes, a chemical necessary for the female Schistosomes to produce eggs. This novel technique describes new methods to inhibit Trk receptor sites on female Schistosome membranes using Trk inhibitor agent to prevent TGR β proteins from binding to the Trk receptor sites. Thus, preventing SmInAct from being created in female Schistosomes, preventing production of eggs and Schistosomiasis.

The invention discloses a Schistosoma japonicum chymotrypsin-like protease (SjCTRL), a protein with an amino acid sequence shown in the SEQ ID NO.2, or a protein which has the same function and is formed through replacement, deletion or insertion of one or more amino acids for the protein. Besides, the invention further discloses a preparation method of the SjCTRL. The preparation method comprises steps such as transmembrane structure removed gene sequence amplification of the SjCTRL, recombinant plasmid construction and identification, induction expression and purification of the SjCTRL and the like, and the immunoprotection of the SjCTRL in anti-schistosoma-infection of mice is evaluated. It is preliminarily verified that the SjCTRL can reduce the number of adults of C57BL / 6 mice infected by Schistosoma japonicum and the number of eggs in livers to a certain extent and can serve as a target of a potential anti-schistosoma vaccine.

The invention provides an application of praziquantel in preparing medicines for resisting Japanese schistosomiasis liver fibrosis. After carrying out disinfestation treatment on mice infected with schistosoma japonica, fibrosis resistant treatment is carried out by continuously applying the praziquantel so as to ensure that the liver fibrosis level is obviously improved, the serum glutamic pyruvic transaminase level is obviously reduced, and the splenomegaly signs of schistosomiasis are also obviously improved.

The invention discloses application of an IRF4 gene in resisting schistosoma infection, in particular to application of the IRF4 gene in preparing or screening a medicament for resisting the schistosoma infection. Meanwhile, the invention also discloses a medicament for resisting the schistosoma infection. The medicament comprises an agent capable of inhibiting or silencing the expression of the IRF4 gene. The invention utilizes IRF4 gene knock-out mice and wild mice to carry out experiments, finds that the IRF4 gene deletion can effectively inhibit the formation of liver granuloma of the micewith schistosoma infection, and obviously improves the liver function of the mice with schistosoma infection. The invention provides a new treatment target for resisting the schistosoma infection, provides a new thought for preparing and screening the medicament for resisting the schistosoma infection, and has great medicinal prospect.

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 protein sequence for killing Japanese Schistosoma and preventing and treating Japanese Schistosoma infection and the corresponding gene sequence and its usage. The protein sequence uses affinity chromatography, polyacrylamide gel electrophoresis; dialysis, freeze drying and protein amino end test sequence to separating obtain the blood serum albumin from oriental hamster unguent. It uses polyase chain reaction technology to obtain oriental hamster blood serum albumin gene sequence. The gene and the decorated mutant insert into the eukaryon expression holder in the animal body, which can be used in preventing and treating the gene of Japanese Schistosoma infection.