64 results about "Cystic echinococcosis" patented technology

The invention discloses a recombinant antigenic protein for diagnosing echinococcosis granulosus (having an amino acid sequence represented by SEQIDNo.1). In addition, the invention also discloses the preparation method of the recombinant antigenic protein, which comprises: amplifying an EgEPC1 gene by using RT-PCR; cloning the EgEPC1 gene in a pGEM-T vector; connecting the EgEPC1 gene with an expression vector PET28a(+) to form a recombinant plasmid PET28a-EgEPC1; transforming the recombinant plasmid PET28a-EgEPC1 to Escherichia coli BL21(DE3) and expressing the recombinant protein through IPTG induction; and identifying a purified recombinant antigen by using SDS-PAGE and Western blotting. In addition, the invention also discloses the diagnosis use of the recombinant antigenic protein. Experiments show that the recombinant antigenic protein of the invention has the advantages of high sensibility and specificity for the diagnosis of echinococcosis granulosus, and has a promising application prospect in the diagnosis of the echinococcosis granulosus.

The invention belongs to the field of bioengineering and provides an Echinococcus granulosusglutathione transferase gene a base sequence of which is shown in SEQ ID NO: 1. The invention also provides a protein encoded by the Echinococcus granulosusglutathione transferase gene. The amino acid sequence of the protein encoded by the Echinococcus granulosusglutathione transferase gene is shown in SEQ ID NO: 2, or is the same as 1st-219th amino acid sequences as shown in SEQ ID NO: 3. The recombinant protein can be used for immunodiagnosis of cystic echinococcosis patients and can be used for coating a plate; and the serums of the patients having cystic echinococcosis, alveolitoid echinococcosis, cysticercosis and other parasite diseases and healthy people are detected by using an enzyme-linked immunosorbent assay (ELISA), and the obtained sensitivity and specificity are respectively 72.41% and 92.36%.

The invention discloses a recombinant antigen protein (of which the amino acid sequence is shown as SEQ ID NO:1) for diagnosing echinococcosis granulose. Moreover, the invention further discloses a preparation method of the recombinant antigen protein. The method comprises the following steps of: amplifying an EgENO gene by adopting RT-PCR (Reverse Transcription-Polymerase Chain Reaction); cloning the EgENO gene into an expression vector PET28a (+) for constructing a recombined plasmid PET28a-EgENO; converting into escherichia coli BL21(DE3); inducing the expression of a recombinant protein through IPTG (Isopropyl-beta-d-Thiogalactoside); and identifying a purified recombinant antigen by using SDS-PAGE (Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) and Western blotting. In addition, the invention further discloses a diagnosis application of the recombinant antigen protein. As proved by an experiment, the recombinant antigen protein has the advantages of high sensitivity, high specificity and the like for the diagnosis of the echinococcosis granulosa, and has a wide application prospect on the aspect of diagnosis of the echinococcosis granulosa.

The invention belongs to the field of bioengineering and discloses an immune chromatography test strip for detecting cystic echinococcosis and alveolar echinococcosis, which comprises a back plate. A sample cushion is arranged at one end of the upper side of the back plate, and a water absorption cushion is arranged at the other end of the upper side of the back plate, a cellulose membrane is arranged between the sample cushion and the water absorption cushion, and a gold-labeled cushion is arranged between the sample cushion and the cellulose membrane. The cellulose membrane is provided with a first detecting line, a second detecting line and a quality control line. The first detecting line contains a purified thick antigen specific to the cystic echinococcosis, the second detecting line contains a recombinant antigen specific to the alveolar echinococcosis, and the quality control line contains antibodies or antiantibodies capable of being in specificity combination with a colloid gold-labeled probe. The invention further provides a preparation method of the immune chromatography test strip for detecting the cystic echinococcosis and the alveolar echinococcosis. The immune chromatography test strip has the advantages of simplicity, sensitivity, specificity and rapidity and being suitable for clinic and on-site use.

The invention discloses a PCR (Polymerase Chain Reaction) detection kit for cystic echinococcosis of a dog. According to the method, a pair of primers is designed according to a 12S rDNA (ribosome Deoxyribose Nucleic Acid) sequence conserved region of pathogene echinococcus granulosus of the cystic echinococcosis, a PCR reaction is carried out with the DNA contained in the wastes of an animal to be detected serving as a template, and then a comparison is carried out relative to an attached negative control and a positive control, in order to determine whether the detected animal infects the echinococcosis. The detection kit is simple and easy to use, and high in sensitivity and stability and can be used for monitoring the health condition of a pet dog.

The invention provides a novel positive antigen of echinococcus granulosus and an application of the antigen and in particular related to an echinococcus granulosus lactated dehydrogenase (EgLDH), a gene or an application of a detection reagent of the echinococcus granulosus lactated dehydrogenase. The echinococcus granulosus lactated dehydrogenase or the gene thereof is applied to the aspect as follows: (i) preparation of positive antigen of echinococcus granulosus or echinococcosis granulosis cyst; and / or (ii) preparation of a reagent or a kit for diagnosing echinococcus granulosus or echinococcosis granulosis cyst. The antigen is highly excellent in diagnostic sensitivity and specificity on the echinococcus granulosus; the diagnostic efficiency of the antigen is highly superior to that of other conventional echinococcus granulosus antigens.

The invention discloses an immunochromatography test strip for detecting cystic echinococcosis and a preparation method thereof. The test strip comprises a hemofiltration membrane sample pad, a gold-marking pad, a cellulose membrane and a water absorption pad, wherein the gold-marking pad is tightly connected with the hemofiltration membrane sample pad and comprises an antibody labeling colloid gold probe of anti-echinococcus antigen advantage special antibody subtypes, the cellulose membrane is tightly connected with the gold-marking pad, the water absorption pad is tightly connected with the other end of the cellulose membrane, a detection line and a quality control line are arranged on the cellulose membrane, the detection line comprises crude antigens aiming at the cystic echinococcosis, and the quality control line comprises anti-antibodies capable of realizing special combination with the antibodies of the anti-echinococcus antigen advantage special antibody subtypes. The immunochromatography test strip of the invention has the advantages of simplicity, convenience, sensitivity, specificity and high speed, and is applicable to clinical and field use.

The present invention discloses a fusion antigen for detecting echinococcosis, a coding gene thereof, a host cell and a kit. The fusion antigen for detecting echinococcosis includes a cystic hydatid antigen portion A, a vesicular hydatid antigen portion B, and a spacer sequence between the antigen portion A and the antigen portion B. The antigen portion A includes multiple epitopes and located atan N-end of the fusion antigen, the antigen portion B is a single epitope and is located at a C-end of the fusion antigen, the spacer sequence chemically links the antigen portion A with the antigen portion B to achieve fusion. The fusion antigen has higher sensitivity and specificity.

The invention provides the preparation of an echinococcus granulosus recombinant BCG vaccine. The vaccine is good in heat stability and convenient to transport and store; single immunity can induce high potency antibody aiming at target antigen, so that immunity is durable, immune time is few, immune procedure is simplified, and immune effect aiming at echinococcus granulosus is enhanced.

The invention discloses a detection card for quickly quantitative detection of echinococcosis antibody in serum. The detection card includes a detection card case and a test paper strip assembled therein. The test paper strip includes a plastic base board provided with pressure-sensitive adhesive, wherein a sample pad, a marker pad, a nitrocellulose membrane and water absorption paper are successively adhered to the base board. The marker pad is composed of a carrier base layer and a marker. The marker is a film formed by spray-coating the carrier base layer with lanthanum-series fluorescencedetection microspheres and lanthanum-series fluorescence quality control microspheres. The nitrocellulose membrane is coated with an echinococcosis recombinant antigen to form a detection line and rabbit-anti-chicken IgY antibody to form a quality control line. The marker includes the fluorescence detection microspheres labeled by the echinococcosis recombinant antigen and the fluorescence qualitycontrol microspheres labeled by the chicken IgY antibody. The detection card can achieve quick quantitative detection of the echinococcosis antibody in situ and has great practical and application values.

The invention discloses an interfering RNA for inhibiting and treating an echinococcus granulosus disease. The interfering RNA targets a GRP78 gene of echinococcus granulosus and has the target sequence of CCTCGTGGTTTACCTCAAA. By transfecting protoscoleces of echinococcus granulosus through electroporation, siRNA-GRP78 provided by the invention can destroy structures of the protoscoleces of cystic echinococcosis, induce apoptosis of the protoscoleces and reduce the activities of the protoscoleces so as to inhibit and kill the protoscoleces of echinococcus granulosus.

The invention provides a novel hydatidosis diagnosing antigen and an application thereof. Specifically, the invention provides echinococcus granulosus cystatin abbreviated to EgCystatin or genes of echinococcus granulosus or an application of a detection reagent of echinococcus granulosus. The EgCystatin, the genes of echinococcus granulosus and the detection reagent are used for preparing a positive antigen of echinococcus granulosus and / or used for preparing a reagent or a kit for diagnosing echinococcus granulosus. The antigen has very high diagnosis sensitivity and specificity to echinococcus granulosus and has the diagnostic efficiency higher than other conventional echinococcus granulosus antigens.

The invention discloses a making method of echinococcosis drug slow-release agent, which comprises the following steps: 1. allocating raw material and biological compatible material; 2. putting biological compatible material into dense fusing machine to stir until the material is fused completely; 3. putting raw material drug into the fused material to stir continuously; 4. blending drug and material completely; fetching the composition out of dense fusing machine; cooling under indoor temperature to form even solid dispersant; 5. grinding solid dispersant; pressing directly; or squeezing through mould after fusing; obtaining the product.

The invention discloses a one-tube nested real-time quantitative PCR detection test kit for detecting alveolar echinocococosis and cystic echinococcosis by micro-sample typing. The test kit is characterized in that inner and outer primers, probes and selection of PCR amplification conditions are designed to finish nested PCR by a one-tube method, and aerosol pollution caused by uncovering is avoided in the operation process; and the detection result accuracy is increased. The detection test kit contains PCR inner and outer primer pair sequences, an MGB typing detection probe sequence, a humanized internal reference gene primer pair sequence for internal quality control and a probe sequence. According to the test kit, samples containing alveolar echinocococosis and / or cystic echinococcosiscan be detected by typing aiming at trace samples such as serum circulating DNA of a patient, the accuracy is high, and alveolar echinocococosis and cystic echinococcosis source specific DNA sequencescan be accurately detected by typing; the alveolar echinocococosis and cystic echinococcosis can be quickly classified and identified on a large scale; and the operation is simple and convenient, andthe trace insect source DNA in the sample can be amplified to reach the typing detection level.

The invention discloses a cystic echinococcosis diagnostic kit and application thereof. The kit comprises a coating antigen composed of an eB2-3 conjugate and / or an eB3-3 conjugate; the eB2-3 conjugate represents a complete antigen obtained by coupling of eB2-3 with carrier protein; the eB3-3 conjugate represents a complete antigen obtained by coupling of eB2-3 with carrier protein; eB2-3 represents polypeptide shown by the sequence 1; and eVP1-2 represents polypeptide shown by the sequence 2. The kit is high in specificity, sensitivity and accuracy, simple and rapid in operation, and suitable for rapidly screening and detecting a lot of serum antibody infected by echinococcus granulosus by veterinary departments and entry-exit inspection and quarantine bureaus in all levels.

The invention discloses a POCKIT Micro fluorescent PCR platform-based cattle and sheep echinococcosis granulose detection kit, and applications thereof. The POCKIT Micro fluorescent PCR platform-based cattle and sheep echinococcosis granulose detection kit comprises a DNA extracting solution, a fluorogenic quantitative PCR reaction liquid, a positive reference substance, and a negative reference substance; the fluorogenic quantitative PCR reaction liquid comprises a cattle and sheep echinococcosis granulose upstream primer, a cattle and sheep echinococcosis granulose downstream primer, a cattle and sheep echinococcosis granulose probe, Taq enzyme, premixed buffer, and deionized water. The nucleotide sequences of the cattle and sheep echinococcosis granulose upstream primer, the cattle and sheep echinococcosis granulose downstream primer, and the cattle and sheep echinococcosis granulose probe are represented by SEQ ID NO.1, SEQ ID NO.2, and SEQ ID NO.3 respectively. The POCKIT Micro fluorescent PCR platform-based cattle and sheep echinococcosis granulose detection kit is high in specificity and sensitivity; no pollution is caused; electrophoresis is not necessary; and one-site rapid detection can be realized.

The invention relates to the technical fields of a yolk antibody, a preparation method thereof and an application thereof, and discloses an echinococcus granulosus hyperimmune yolk antibody, a test strip, a preparation method and application. The echinococcus granulosus hyperimmune yolk antibody is obtained according to the following method: preparation of echinococcus granulosus antigen protein,preparation of immunogen, feeding and immunization of laying hens, and preparation of echinococcus granulosus hyperimmune yolk antibody. The present invention discloses for the first time an echinococcus granulosus hyperimmune yolk antibody, a freeze-dried hyperimmune yolk antibody and the use thereof in the diagnosis of echinococcosis, and when a colloidal gold chromatographic test strip made ofthe echinococcus granulosus hyperimmune yolk antibody and the freeze-dried hyperimmune yolk antibody of the present invention is used to diagnose echinococcosis, the colloidal gold chromatographic test strip has good sensitivity, specificity, accuracy and stability, and a simpler detection method for diagnosis of echinococcosis with lower cost is provided.

The invention relates to the technical field of human and animal alveolar echinococcosis (AE) antibody detections, specifically to cystic echinococcosis (CE) cyst fluid degreasing antigen for detecting AE antibody, and a preparation method thereof. According to the present invention, the CE cyst fluid degreasing antigen is used for detecting AE antibody by using an enzyme-linked immunosorbent test double antigen sandwich method so as to effectively increase quality of the detection reagent, and provide characteristics of strong specificity, high sensitivity, stable performance and wide uses; the CE cyst fluid degreasing antigen can be used for detecting human and animal AE antibodies so as to conveniently compare antibody contents of different animal varieties; and according to the immunological reaction principle, the CE cyst fluid degreasing antigen further can be used for preparing colloidal gold immunochromatography reagents and other rapid detection reagents for detecting AE antibodies so as to create favorable conditions for echinococcosis identification diagnosis, prevalence surveys and epidemic situation surveillance works.

The present invention discloses application of ERK inhibitor PD98059 in preparation of a medicine for treatment of alveolar hydatid diseasein, and the ERK inhibitor PD98059 is used for killing alveolar hydatid diseasein pathogen-bubble hydatid tapeworm, has significant hydatid protoscolex inhibition and killing effect, and can be used for treatment of alveolar hydatid disease.

The invention relates to the technical filed of an insulin growth factor acceptor inhibitor PQ401, and concretely relates to an application of the insulin growth factor acceptor inhibitor PQ401 as a medicine for treating echinococcosis granulosa. The insulin growth factor acceptor inhibitor PQ401 is a novel echinococcosis resistance medicine; in vivo and in vitro pharmacodynamic experiment data show that the PQ401 is an efficient echinococcosis resistance medicine molecule, can destroy the tissue structure of the Echinococcus granulosus vesicles and causes necrosis of a large amount of cells of the Echinococcus granulosus vesicles; in vivo pharmacodynamic experiment data show that the medicine has same echinococcosis granulose treatmetn effect with clinically commonly used albendazole; and the long term administration of albendazole by clinic echinococcosis granulosa patients causes medicine resistance, so the medicine can substitute albendazole to be used in the treatment of the echinococcosis granulosa.

The invention relates to a rapid detection kit for a dog mature echinococcosis antigen and a use method of the rapid detection kit. The rapid detection kit comprises a sample pad, a gold mark pad, a nitrocellulose membrane, a water absorption pad and a base plate, wherein the sample pad is fixed at the top end of the left part of the base plate, the gold mark pad is fixed between the right part of the sample pad and the base plate, the water absorption pad is fixed at the top end of the right part of the base plate, and the nitrocellulose membrane is fixed between the gold mark pad and the water absorption pad. The rapid detection kit for the dog mature echinococcosis antigen (colloidal gold method) has relatively high sensitivity, specificity and stability, can be carried along, can be used for carrying out field testing at any time anywhere and does not require special testing auxiliary equipment, so that the testing time is shortened, and the testing efficiency is improved; and besides, the rapid detection kit is used for testing dog mature, so that the injury caused to dogs is avoided, and the detection work is relatively simple and convenient.

The invention discloses a use of a TGF-beta 1 receptor blocker in preparation of drugs for preventing echinococcosis recurrence. The applicant research shows that after use of the TGF-beta 1 receptor blocker in the mouse early suffering from echinococcosis granulosis cyst, a downstream molecule Smad2 / 3 phosphorylated protein expression level is reduced, NK cell active receptor NKG2D expression is improved, a target cell cracking rate of NK cells is recovered, NK cell activity is in a positive correlation relationship with active receptor NKG2D correlation analysis expression, a ratio of CD4<+> / CD8<+>T cell is high, and the amount of CD4<+>CD25<+>T cells. Therefore, the TGF-beta 1 receptor blocker can effectively prevent echinococcosis recurrence in clinical application.

The present invention relates to a compound preparation for treating large yellow croaker irritating cryptocystosis and its application. The compound preparation is composed of 10 wt% to 15 wt% of nitazoxanide and 5 wt% to 7 wt% of copper sulfate , oxytetracycline 25 wt%~30 wt%, dissolving agent 48%~60%, the sum of the above raw materials is 100%. The method of use is: after the diagnosis of cryptocystosis irritant, sprinkle the above compound preparation in an amount of 8~10 g / m3, and soak in the bath continuously for 3 days, which can effectively cure cryptocystosis irritant of large yellow croaker and prevent secondary infection of bacteria . The invention has the advantages of simple preparation method and convenient use, can significantly reduce the death rate of large yellow croaker stimulated by cryptocystosis, and improve the economic benefits of breeding.

The invention discloses an echinococcosis granulosa ELISA antibody detection kit. The detection kit comprise an EgAgB8 / 2 recombinant antigen pre-coating enzyme label plate, a sample weak solution, positive contrast liquid, negative contrast liquid, an enzyme conjugate, washing liquid, a substrate coloring solution and a stopping solution. A method for preparing the EgAgB8 / 2 recombinant antigen comprises the following steps: using RT-PCR for amplifying EgAgB8 / 2 gene cDNA, cloning an EgAgB8 / 2 gene, constructing a prokaryotic expression vector pET-EgAgB8 / 2, and performing expression in escherichia coli BL21(DE3), using the IPTG-induced prokaryotic expression vector for expression, performing centrifugation to collect thalline, using a bacteria lysate for dissolving, and performing ultrasonicfragmentation, performing centrifugal collection of a supernatant, and using a Ni-NTA glue for purification. The kit takes recombinant protein EgAgB8 / 2 as antigen to detect an animal echinococcosis granulosa antibody, can be used for qualitative detection of the animal echinococcosis granulosa antibody, and has the advantages of stable antigen source, good specificity, high sensitivity, and good repeatability.

The embodiment of the invention provides a method for detecting cystic echinococcosis. The method comprises the following steps: coating an ELISA plate with an Ag5 antigen which is recombined and induced by a prokaryotic expression system and is purified in vitro; diluting serum of a subject in proportion, and adding the diluted serum and known negative and positive serum into the plate holes of the ELISAplate for reaction; and measuring the OD450 value of each plate hole, and calculating the sample value of each plate hole according to the publicity. Clinical serum sample detection shows that the detection specificity and sensitivity are higher than those of a commercial kit, and false positive results are effectively eliminated. The indirect ELISA method established in the invention provides a more accurate and effective detection means for serological diagnosis of echinococcosis.

The invention relates to a gene rEg.myophilin molecular engineering vaccine resistant to sheep echinococcosis infection. The vaccine is obtained by extracting RNA from a China strain of Echinococcus granulosus, obtaining Eg.myophilin gene through RT-PCR and performing colonizing, induced expression and purification, and collected under CCTCC M2015426. The invention further relates to application of rEg.myophilin gene molecules in preparing vaccines resistant to sheep echinococcosis infection. Immunoprotection verification of an Echinococcus granulosus recombinant vaccine is performed on sheep, and immune protection efficiency reaches 92%. The vaccine is expected to play a great role in preventing and treating liver echinococcosis.

The invention relates to the technical field of echinococcosis drugs, in particular to an application of a glucose transporter 1 inhibitor WZB117 to preparation of a cystic echinococcosis treating drug. The invention discloses the application of the WZB117 to preparation of the cystic echinococcosis treating drug for the first time. Pharmacodynamic experiment data in vivo and in vitro show that the WZB117 is an efficient echinococcosis-resistant drug molecule, and particularly has a remarkable treatment effect on cystic echinococcosis, and the effect is superior to that of albendazole sulfoxide which is an albendazole metabolite; the combined use of the WZB117 and the albendazole sulfoxide has a better synergistic treatment effect, and the effect is remarkably superior to that of independent administration of the WZB117 and independent administration of the albendazole sulfoxide; and meanwhile, the dosage of the combined use of the WZB117 and the albendazole sulfoxide is reduced by 1 / 3-1 / 2 compared with the dosage of the independent administration of the albendazole sulfoxide, so that the dosage and the drug resistance are greatly reduced, a good treatment effect is achieved, and the WZB117 can be used as an effective substitute for existing albendazole.

The invention discloses application of gynesine in preparing a drug for in vitro treating echinococcosis granulosa, relating to the technical field of medicines. The gynesine disclosed by the invention is used for preparing the drug for in vitro treating echinococcosis granulose. The gynesine is a fenugreek plant extract and has slight side effect to a human body.

The invention relates to the technical field of bio-medicine engineering and discloses an interfering RNA (Ribonucleic Acid) for inhibiting expression of an echinococcus granulosus protoscolex DNA (Deoxyribonucleic Acid) oxidative damage repairing gene and application of the interfering RNA. The interfering RNA for inhibiting expression of the echinococcus granulosus protoscolex DNA oxidative damage repairing gene has target sequences of forward CCGUGUUCGUAAGCAGCUUTT and backward AAGCUGCUUACGAACACGGTT. The invention discloses the target sequences of the interfering RNA for inhibiting expression of the echinococcus granulosus protoscolex DNA oxidative damage repairing gene for a first time, and also discloses application of the interfering RNA in preparing cystic echinococcosis medicines for the first time. By inhibiting expression of the EgRPS9 gene by using the interfering RNA, growth of echinococcus granulosus protoscolex is inhibited and killed by affecting the repairing function of DNA oxidation damage of the echinococcus granulosus protoscolex, so that cystic echinococcosis can be effectively treated.

The invention relates to the technical field of biology, and particularly discloses applications of echinococcus granulosus dihydrofolate reductase to preparing an immunizing antigen and / or a kit which are / is used for detecting echinococciasis granulous. The echinococcus granulosus dihydrofolate reductase serves as the immunizing antigen and can be identified by sheep positive blood serums naturally infected by the echinococciasis granulous; when the echinococcus granulosus dihydrofolate reductase is applied to indirect ELISA detection, the high specificity and the high sensitivity are achieved, and the clinical-detection coincidence rate is up to 97.9%. A detection method built with the echinococcus granulosus dihydrofolate reductase serving as the immunizing antigen has the good diagnosing effect, and can be used for preliminary screening of the sheep echinococciasis granulous in an affected area.