61 results about "Schistosoma sinensium" 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 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 object of the present invention is the detection of Schistosoma species in biological samples by PCR. The first embodiment of the present invention relates to a method for diagnosing schistosomiasis, that is, amplifying the DNA sequence of Schistosoma species by PCR, then separating the amplified products by electrophoresis, and finally detecting by appropriate techniques. In a second embodiment, the invention is directed to a diagnostic kit for the detection of schistosomiasis. The basic kit contains all the necessary reagents for PCR technology, that is, specific primers, nucleotides, and appropriate buffers for PCR amplification. The kit may optionally include Taq polymerase in sufficient quantity for amplification, standard DNA as a positive control for the reaction, buffer for preparing amplified material for electrophoresis, and a protocol and instruction manual for users.

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 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 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 present invention relates to a preparation method of a snail respiratory protein. A respiratory protein containing copper ions is extracted from a snail vivo used as an intermediate host of Schistosoma japonicum by virtue of mechanical extrusion, tissue dissection, hemolymph extraction, ammonium sulfate precipitation, super high-speed centrifugal separation and other means. The respiratory protein contains 0.2-0.3% of copper, the UV characteristic absorption peak is at 275nm and 340nm, and a cylindrical agreegate structure is observed under a transmission electron microscope.

The invention discloses a method of fast separating Japan schistosome unfledged spawn, which employs technique of combining repeat sieving and pancreatin processing and completes in two hours. Purity of the obtained spawn is above 99%, structure of the spawn is integrated and good biological activity is remained, which can satisfy basic need of antigen extraction and analysis. Comparing to exist technique, yield of the spawn is largely increased.

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 invention provides a primer group for detecting schistosoma haematobium. The primer group comprises an upstream primer and a downstream primer, wherein the sequence of the upstream primer is shownas SEQ ID No. 1; and the sequence of the downstream primer is shown as SEQ ID No. 2. The primer group, probe and kit for detecting the schistosoma haematobium are simple and convenient to operate; tests prove that when the primer group, probe and kit for detecting the schistosoma haematobium are used for detecting the schistosoma haematobium, the specificity is high, the sensitivity and the accuracy are high, the repeatability is good, a detection result can be obtained within 20 minutes, the requirements on detection conditions and the technical requirements of detection personnel are low, and the detection cost is low.

The invention discloses a primer, a probe, a kit and a method for visually and rapidly detecting nucleic acid of schistosoma japonicum katsurada through LFD-RPA. The sequence of the primer is shown as SEQ ID NO.1-2, and the sequence of the probe is shown as SEQ ID NO.3. The RPA primer and the probe are designed by taking a schistosoma japonicum G55A (SjG55A) gene as a target sequence, the sensitive and rapid visual detection of schistosoma japonicum nucleic acid is realized by adopting an RPA amplification technology in combination with a lateral flow chromatography test strip method, the detection limit of schistosoma japonicum genome DNA can reach 10fg, and the identification detection of schistosoma japonicum and schistosoma mansoni is expected to be realized. One positive oncomelania in 1500 negative oncomelania can be detected, operation is simple and rapid, no special instrument is needed, the reaction temperature is close to the room temperature, the result can be observed by naked eyes, and detection and monitoring of the schistosome intermediate host in a laboratory and on site are facilitated.

The present invention relates to the field of biological technology. The pachysand extract or steroid alkaloid produced through crushing and magnetic stirring extraction process is compounded into water solution of different concentration to kill Oncomelania hupensis as the host of Schistosoma japonicum. It has obvious killing effect and is harmless to human body and farm animal.

The invention discloses a Wnt family gene which is amplified for the first time from Schistosoma japonicum 19-day schistosomulum by utilization of RACE technology, wherein, sequence analysis indicates that: a complete coding frame of the gene comprises 1311bp, 436 amino acids coded; and theoretical molecular weight of 49.6kD. Homology analysis results indicate that: amino acid sequences of the gene have typical Wnt family protein characteristics; similarity of the amino acid sequences with amino acid sequences of Dugesia japonica and human Wnt4 is relatively 43 percent and 37 percent; the gene is presumed to be a Wnt4 gene of schistosome and named as Sjwnt4 (GenBanK Log-On No.: DQ643829). Real-time quantitative PCR analysis indicates that: the gene is expressed all in 14-day schistosomulum, 19-day schistosomulum, 31-day imago, 44-day female worms and 44-day male worms. The invention constructs a pronucleus expression vector pGEX-4T-2-Sjwnt4 of the gene and applies an Escherichia coli system for expression; expression proteins exist in the form of inclusion bodies; Western blottings indicate that expression products can be identified by crude antigen immune serums of Schistosoma japonicum imago.

The invention provides a nucleic acid diagnosis kit for detecting schistosoma japonicum. The kit provides a group of primers which have high sensitivity and high specificity and are used for detectingschistosoma japonicum, wherein the primers have nucleic acid sequences as shown in SEQ ID NO.1-2. The invention constructs real-time fluorescent quantitative PCR detection technology with high sensitivity and high specificity for schistosoma japonicum free nucleic acids, and the detection technology can be used for accurately and quickly detecting infection of schistosoma japonicum, determining whether a water body contains miracidia and cercariae or not and performing species identification on schistosoma japonicum.

The invention discloses a RPA primer and a method for detecting schistosoma japonicum katsurada, particularly relates to a biological detection technology, and belongs to the technical field of molecular biology. A pair of specific primers is designed according to an Sj28S ribosome gene sequence of the schistosoma japonicum, and an RPA method for nucleic acid detection of the schistosoma japonicum is further established. The method can specifically amplify and detect the nucleic acid of the schistosoma japonicum katsurada, and the lowest detectable genome DNA concentration is 100 fg/mu L. The product has the advantages of high specificity, high sensitivity and high stability, can realize the index amplification of target fragments within 20 minutes, has low requirements on environmental equipment, is suitable for the detection of large-batch samples, and has a better field application prospect.

The invention relates to a schistosoma japonicum heat shock protein (SjHSP90) recombinant protein and application thereof to schistosomiasis diagnosis and treatment effect evaluation and belongs to the field of immunology. The invention provides the preparation of the SjHSP90 recombinant protein, wherein the amino acid sequence of the SjHSP90 recombinant protein is shown as SEQIDNO.1. The SjHSP90 recombinant protein is further applied to schistosomiasis diagnosis and treatment effect evaluation. According to the invention, the SjHSP90 recombinant protein is applied to the schistosomiasis diagnosis for the first time; the SjHSP90 recombinant protein can be used for not only the schistosomiasis diagnosis and but also the treatment effect evaluation, so that the SjHSP90 recombinant protein has broad market space and good social and economic benefits.

A double peptide-DNA vaccine based on T-cell epitope for preventing and treating the Japanese schistosome infection features that its eucaryotic expression carrier carrying the epitope peptide coding gene is wrapped by the protein containing the epitope peptide. The amino acid sequence of said T-cell epitope peptide is AKQYNICCKFKELLD. Said vaccine can be prepared by cationic peptide carrying technique.

The invention discloses siRNA of schistosoma japonicum SjELAV-like 1 genes. The siRNA has nucleotide sequences shown as SEQ ID NO.3 and SEQ ID NO.4. The invention further discloses the application ofthe siRNA of schistosoma japonicum SjELAV-like 1 genes. The siRNA of schistosoma japonicum SjELAV-like 1 genes disclosed by the invention is capable of obviously inhibiting transcription of the schistosoma japonicum SjELAV-like 1 genes and obviously reducing the number of liver loaded eggs and egg hatching rate of schistosomiasis infected mice, causes certain damages to the tegumental structure ofschistosoma and causes morphological abnormality of germ cells, and is applicable to preparation of drugs for treating schistosomiasis.

The invention discloses a method for detecting schistosoma japonicum infection by using a host exosome miRNA-142a-3p. A nucleotide sequence of the serum exosome miR-142a-3p is as shown in SEQ ID NO 1,and the method for detecting the schistosoma japonicum infection is established based on primers with a nucleotide sequence as shown in SEQ ID NO 2. The method for detecting the schistosoma japonicuminfection by using the host exosome miRNA-142a-3p is rapid, sensitive, specific and stable to detect. According to the method, serum of patients is directly extracted so that the trauma of the patients is reduced, and the trauma is small; a q-PCR detection method is used, and the sensitivity is high; and the q-PCR detection method has high sensitivity. At present, a mature exosome extraction kitis provided, meanwhile, q-PCR detection is a commonly used technology in a laboratory and is relatively simple. Therefore, the method is worth popularizing and has a good clinical application value.

The invention discloses siRNA of schistosoma japonicum prohibitin PHB1 and PHB2 genes. The siRNA comprises a nucleotide sequence, which is hybridized with a target sequence of the schistosoma japonicum PHB1 and/or PHB2 genes. The invention further discloses application of the siRNA of the schistosoma japonicum PHB1 and PHB2 genes. The siRNA of the schistosoma japonicum PHB1 and PHB2 genes in the invention can obviously inhibit transcription of SjPHB1 and SjPHB2 genes, and can induce a mouse to obtain 40.43% worm reduction rate (P is less than 0.05); and the siRNA is suitable for preparing a medicine for treating schistosomiasis.