35 results about "Dermatophagoide" patented technology

The invention provides regenerated cellulose fiber containing a microencapsulated anti-mite medicine and a preparation method thereof. The regenerated cellulose fiber comprises a viscose-rayon spinning solution and the microencapsulated anti-mite medicine, the microencapsulated anti-mite medicine accounts for 0.05-2% by mass of methyl cellulose in the viscose-rayon spinning solution, methyl cellulose accounts for 99.95-98% by mass of dry regenerated cellulose fiber, and borneol and argy wormwood essential oil are taken as a capsule core. The preparation method comprises: firstly preparing a solution of borneol and argy wormwood essential oil, then preparing microcapsules from the solution of borneol and argy wormwood essential oil through a microencapsulation technology, adding the microcapsules into the viscose-rayon spinning solution, mixing uniformly, and finally utilizing a viscose-rayon wet spinning technology to prepare the regenerated cellulose fiber containing the microencapsulated anti-mite medicine. The regenerated cellulose fiber containing microencapsulated anti-mite medicine has the repelling rate on dermatophagoides farina of 95% or more, and has extremely strong anti-mite effect, and has the antibacterial rate on staphylococcus aureus, escherichia coli and other pathogens of 90% or more.

Isolated nucleic acids encoding an allergen of Dermatophagoides pteronyssinus, Der p III, are disclosed. A cDNA encoding a peptide having a Der p III activity and a predicted molecular weight of about 24,985 daltons is also described. The nucleic acids can be used as probes to detect the presence of Der p III nucleic acid in a sample or for the recombinant production of peptides having an activity of Der p III. Peptides having an activity of Der p III can be used in compositions suitable for pharmaceutical administration or methods of diagnosing sensitivity to house dust mites.

The invention discloses a method for developing a novel indoor acaricide. The novel acaricide uses clove and other Chinese medicinal materials as raw materials, the main active component thereof is eugenic acid, and the novel acaricide is a pure natural chemical substance, has no notable harms to human bodies and other organic bodies, has no potential hazards to the environment too, and can be widely applied to killing, preventing and controlling various dust mites of dermatophagoides pteronyssinus, dermatophagoides farinae and the like in various regions and various indoor environments. The novel acaricide can effectively control the number of the dust mites in the living environments of people, greatly reduce the density of dust mite allergens in the indoor environments, and significantly reduce the disease incidence of dust mite anaphylactic diseases. Besides, the novel acaricide has outstanding effects of preventing and performing adjuvant therapy on various anaphylactic diseases taking the dust mites as the allergens, such as allelgic asthma, rhinitis and the like.

The invention belongs to biotechnics in particular a cultivation method of dust mite of room. The steps of said method comprises: collection dust with mites, little cultivation in early stage, a vast amount of cultivation in mast bottle by collecting mites with fine needle and repeat collection by saturated saline flotation method, to cultivate dust mites of room in controlled temperature and humidity, to reach effective performance. The invention avoids a lot amount of work and time, pollution when cultivating in open container, low production of mites, and the degree of purity is more than 90 % contrasted to low degree of purity by traditional method; and the cultivation base is easy to attained and in rational proportion.

The invention discloses a method for producing recombinant dermatophagoides farinae allergen Der f1 and Der f2 fusion protein and relates to the technical field of construction of dermatophagoides farinae allergen gene by a biological technology. The method comprises the following steps of: obtaining a coding gene by extracting the total RNA of the dermatophagoides farinae and adopting PCR (Polymerase Chain Reaction) amplification, cutting glue to recover a product, named as Der fm, cloning to pMD19-T vectors, subcloning to expression vectors pET-28a(+), converting the vectors to escherichia coli, carrying out inducible expression by using IPTG (isopropyl-Beta-d-thiogalactoside). The method disclosed by the invention has the advantages that the consumed time is short, the process is simple, the cost is lower, the components such as a solvent can be thoroughly removed, the purity of the allergen can be increased fundamentally, the occurrence of side reaction in immunological therapy is avoided, the purity is high, the classification is clear, the accuracy of diagnosis can be increased, and a standard product is expected to be provided for novel immunological therapy.

The present invention provides isolated peptides of the major protein allergens of the genus Dermatophagoides. Peptides within the scope of the invention comprises at least one T cell epitope, or preferably at least two T cell epitopes of a protein allergen selected from the allergens Der p I, Der p II, Der f I, or Der f II. The invention also pertains to modified peptides having similar or enhanced therapeutic properties as the corresponding, naturally-occurring allergen or portion thereof, but having reduced side effects. The invention further provides nucleic acid sequences coding for peptides of the invention. Methods of treatment or of diagnosis of sensitivity to house dust mites in an individual and therapeutic compositions comprising one or more peptides of the invention are also provided.

The invention provides a method for preparing an allergen diagnostic reagent of acute allergic diseases caused by dermatophagoides pteronyssinus (Der p for short). The allergen diagnostic reagent is Der p allergen pricking liquid. The invention also provides a method for preparing an allergen diagnostic reagent raw material, namely Der p pure mite allergen, a method for evaluating the valence of the allergen diagnostic reagent and a purpose for specifically diagnosing the allergic diseases. The allergen diagnostic reagent achieves the aim for detecting whether a patient is allergic to a matter to cause a disease by a sensitized principle. The Der p pure mite allergen raw material has short culture period and high purity, and the pricking liquid made of the raw material has high activity, thereby greatly reducing a skin test risk of the allergic patient and improving the safety of clinical medical treatment.

Isolated nucleic acids encoding an allergen of Dermatophagoides pteronyssinus, Der p III, are disclosed. A cDNA encoding a peptide having a Der p III activity and a predicted molecular weight of about 24,985 daltons is also described. The nucleic acids can be used as probes to detect the presence of Der p III nucleic acid in a sample or for the recombinant production of peptides having an activity of Der p III. Peptides having an activity of Der p III can be used in compositions suitable for pharmaceutical administration or methods of diagnosing sensitivity to house dust mites.

The invention discloses a semi-quantitative detection kit for house dust mite-specific IgE and its application. The test kit includes a detection fluorescent liquid and an immunochromatographic test strip for semi-quantitative detection of house dust mite-specific IgE; wherein, the detection fluorescent liquid is obtained by adding carboxyl-containing quantum dot nanospheres on the surface It is covalently coupled with goat anti-human sIgE and mouse anti-rabbit IgG, and prepared after mixing; the immunochromatography test strip includes sequentially connected sample pads, nitrocellulose membranes, water-absorbing pads, and an assembly platform for providing On the lower plastic backing, the nitrocellulose membrane has a test line coated with dust mite allergen protein and a quality control line coated with rabbit IgG, and the sample pad provides a place for the sample to be tested to be added. The semi-quantitative detection of house dust mite-specific IgE can be realized by using the kit established by the invention, which has a good correlation with clinical symptoms, high detection sensitivity, easy to use, and is suitable for large-scale population allergen screening and family detection Wait for the environment.

Isolated DNA encoding allergens of Dermatophagoides (house dust mites) particularly of the species Dermatophagoides farinae and Dermatophagolides pteronyssinus, which are protein allergens or peptides which include at least one epitope of the protein allergen. In particular, DNA encoding two major D. farinae allergens, Der f I and Der f II and DNA encoding a D. pteronyssinus allergen, Der p I. In addition, the proteins or peptides encoded by the isolated DNA, their use as diagnostic and therapeutic reagents and methods of diagnosing and treating sensitivity to house dust mite allergens.

The invention discloses a novel sulfonate antibacterial anti-mite agent and a research method for an efficient complexation of the agent. The novel sulfonate antibacterial anti-mite agent comprises quinolin-8-yl 4-chlorobenzenesulfonate and 2,3-dichlorophenyl 4-fluorobenzenesulfonate; and an efficient compound anti-mite agent preferably comprises quinolin-8-yl 4-chlorobenzenesulfonate and 2,3-dichlorophenyl 4-fluorobenzenesulfonate with a ratio of 3:2. The novel anti-mite agent is a sulfonate antibacterial anti-mite agent possessing the advantages of being wide in antibacterial spectrum, efficient, high in durability, safe and the like, and is applicable to substantially reduce levels of dermatophagoides pteronyssinus and dermatophagoides farina allergens of household interior ornaments, carpets, sofas and mattresses. The sulfonate antibacterial anti-mite agent is characterized by possessing both extremely strong mite contact killing capability and extremely strong smell mite killing capability.

The invention discloses a kit detecting dermatophagoides-farinae allergen specific IgE antibody and a method. The kit comprises a calibration substance, a quality-control substance, a biotin-labeled dermatophagoides-farinae allergen solution, an alkaline-phosphatase-labelled mouse anti-human IgE secondary-antibody solution and a streptavidin-labeled nanometer-magnetic-particle suspension. The kit is utilized to detect the dermatophagoides-farinae allergen specific IgE antibody. Through the above manner, the reagent compositions in the kit are good in stability, the expiration data is one year or more, the detection sensitivity is high, the specificity is good, and variation is small. An improved unified technology is obtained after a large amount of experiments are performed for technology optimization, and production is performed strictly according to standard production operation rules and quality control rules. A user can perform standard operation and obtain a reliable result only according to the operation instruction. During clinic research, the coincidence correlation with a foreign imported reagent is up to 90% or more, and the cost only is 1 / 2 of the foreign imported reagent.

The invention relates to a method for performing high-solubility expression and purification on dermatophagoides pteronyssinus major allergen Der p 2 protein in Escherichia coli. According to the method, the Escherichia coli chaperonin trigger factor (TF) is used as a fusion tag to implement high-solubility expression of Der p 2 and obtain the optimized high-efficiency purification technique; and the purity of the sample obtained by the method is higher than 98%. The verification proves that the purified recombinant Der p 2 protein has a consistent secondary structure folding state with the naturally extracted Der p 2 protein and has the identical combination activity with specific IgE in the serum of the dermatophagoides-pteronyssinus-allergic patient. The fusion expression method used in the invention can implement soluble expression of the Der p 2 protein, has the advantages of cuttable fusion tag, high recombinant protein expression level and simple purification technique, is suitable for large-scale industrial production, and can maintain the natural activity of the product.

The invention relates to an optimized gene mderp2 obtained by carrying out codon optimization onto a gene of primary allergen Derp2 of the Dermatophagoides pteronyssinus, a recombinant expression plasmid pN8148-SD constructed based on the gene for secreting and expressing allergen Derp2 and recombinant lactococcus lactis L.lactis SD (preservation number as CGMCC (China General Microbiological Culture Collection Center) No.8223). Western blot results show that the recombinant lactococcus lactis L.lactis SD disclosed by the invention can realize secretion and expression of the allergen Derp2; meanwhile, an animal experiment result shows that orally-taken recombinant lactococcus lactis L.lactis SD can increase level of regulatory T cells in a mouse spleen, and can lower anaphylactic reaction caused by dust mite allergen Derp2 in the mouse body, and has a better preventive effect.

This present invention discloses a monoclonal antibody which specifically recognizes and binds to a epitope of group 2 allergen of Dermatophagoides pteronyssiuns, usually named Der p 2, and a hybridoma cell line producing thereof. Furthermore, this invention also discloses a strip, kit and method utilizing said monoclonal antibody for the detection of the presence of dust mite allergens and the calculation of dust mite number in the environment.

The invention discloses a method for breeding common cheyletid mites by utilizing dermatophagoides farinae. The method comprises the following steps: firstly, sterilizing and drying a breeding container, adding flour, fish bone powder and yeast extract powder in a mass ratio of (5-20):(1-4):(0-3) to serve as breeding foods which reach one fifth to three fifths of the height of the breeding container; then, breeding dermatophagoides farinae int the breeding container to serve as preying mites, introducing common cheyletid mites when the preying mite population density is more than or equal to 100 per gram, and keeping the breeding ratio of preying mites to common cheyletid mites to be (20:1)-(100:1); breeding for 15-60 days to obtain common cheyletid mites completely without preying mites or separating the common cheyletid mites in different containers and keeping on breeding until the population density of common cheyletid mites is more than or equal to 80 per gram; and keeping the breeding temperature to be 17-28 DEG C and a relative humidity of 65-85 percent in the steps. The breeding method is simple, efficient and low in cost, and is suitable for large-scale production.

The invention discloses a preparation method of a Chinese herbal medicine extracting solution for treating rhinitis. The Chinese herbal medicine is mainly prepared from the following Chinese herbal medicines in parts by weight: 20-40 parts of liquorice root, 5-20 parts of centipeda minima, 5-15 parts of flos magnoliae liliflorae, 5-20 parts of luffa stem and 10-30 parts of honeysuckle flower. TheChinese patent medicine belongs to a pure plant Chinese herbal medicine, has the effects of resisting bacteria, diminishing inflammation, inhibiting bacteria and relieving itching, is suitable for allergic rhinitis, nasosinusitis and variability rhinitis caused by various dermatophagoides farinae, has an auxiliary improvement and relief effect, has the advantages of good taste, no peculiar smell,no toxic or side effect, quick effect, remarkable curative effect, low recurrence rate, no demand in decocting, convenience in taking and carrying and the like, and is suitable for more people.

The invention relates to a nasal cavity excitation reagent based on recombinant dermatophagoides farinae Der f2 protein, and belongs to the technical field of biological medicine. The invention provides an application of a recombinant dermatophagoides farinae Der f2 protein in preparation of an allergen nasal cavity excitation reagent. The invention also discloses an application of the recombinant dermatophagoides farinae Der f2 protein in preparation of a diagnostic reagent for diagnosing allergic rhinitis. The reagent comprises a spray, a drop or an injection; the key allergenic peptide fragment of the recombinant dermatophagoides pteronyssinus protein is provided, and it is proved that the key allergenic peptide fragment can cause degranulation of in-vivo mast cells. The protein is applied to a nasal cavity excitation experiment to check whether corresponding IgE exists in nasal mucosa or not, and clear diagnosis can be achieved. The recombinant protein can be prepared into standard concentration to meet the requirement of concentration gradient, so that the safety and accuracy of nasal cavity excitation test are ensured.

The present invention relates to a fusion protein having formula (I): X1-Y-X2, wherein X1 and X2 comprise each four to six allergen fragments or variants thereof fused to each other, wherein said allergen fragments are derived from at least two allergens of the genus Dermatophagoides, and wherein Y is a carrier protein.

According to the invention, modified allergen gene xderp2 is obtained through genetical modification to main allergen Derp2 of dermatophagoides pteronyssinus, and recombinant plasmids and recombinant lactococcus lactis cocci XD containing xderp2 (preservation number is CGMCC No.8220) genes are built. Western blot results show that the protein of the modified allergen gene xderp2 can be expressed in recombinant lactococcus lactis cocci; in addition, animal experiment results show that mite anaphylaxis can be restrained obviously by oral administration of recombinant lactococcus lactis cocci of expression modified dust mite allergen XDerp2.

The present invention discloses method of culturing and collecting Dermatophagoides pterronyssinus (Der p), extracting Der p sensibiligen, and measuring the content of the first Der p sensibiligen component (Der p1) inside Der p extracting liquid. The measurement of Der p1 inside Der p extracting liquid is in double-sandwich monoclonal antibody process. In addition, the present invention also discloses the establishment of standard serum library of allergic Der p patient and the test of the total sensibiligen bioactivity of Der p extracting liquid. The method of the present invention has the features of easy operation and fast speed.

The invention discloses a kit for semi-quantitative allergen screening. The kit comprises a nitrocellulose membrane strip coated with 19 allergens, a biotinylated anti-human IgE antibody, streptavidin labeled by alkaline phosphatase and a substrate solution, the allergens include willow / poplar / elm, common ragweed, mugwort, dermatophagoides pteronyssinus / dermatophagoides farinae, house dust, cat hair, dog epithelium, cockroaches, penicillium notatum / cladosporium / aspergillus fumigatus / alternaria, scandent hop, egg white, milk, peanuts, soybeans, beef, mutton, cod / lobster / scallop, shrimps and crabs. 19 common allergens can be qualitatively and quantitatively detected through one-time detection, and the method has the advantages that the operation is simple, the detection efficiency is greatly improved, the production and use cost is reduced, and the popularization and application of allergen detection are facilitated.

The invention discloses a dermatophagoides farinae microsatellite marker. The microsatellite marker is selected from at least one of Df01, Df02, Df03, Df04, Df05, Df06, Df07, Df08, Df09, Df10, Df11 and Df12, wherein Df01 to Df12 are DNA sequences of SEQ ID NO.01 to Df12 or complementary sequences thereof in sequence, and microsatellite repetition elements of the Df01 to Df12 are (CTG)5-8, (ATG)5-7, (ATC)9-12, (GTT)6-7, (ATC)6-8, (TG)6-9, (ATG)5-9, (GT)7-10, (ATG)4-5, (CAA)5-7, (TTG)6-9 and (CA)7-8 The microsatellite marker and the primer thereof provided by the invention have the characteristics of stable PCR amplification, high polymorphism and the like, and the primer can be used for genetic diversity and genetic structure analysis of dermatophagoides farinae and has relatively high application value.

The invention relates to the field of genetic engineering associated with dust mite allergens, and discloses a hybridoma cell strain AntiTput-13, which can stably secrete a monoclonal antibody againstTyrophagus putrescentiae allergen Der p10. The hybridoma cell strain is collected in the China General Microbiological Culture Collection Center at Beichen West Road, Chaoyang District, Beijing withthe number of CGMCC No. 16298. Tests prove that the monoclonal antibody antiTPUT-DP10 secreted by the hybridoma cell strain can specifically react with Tyrophagus putrescentiae allergen Der p10. Basedon the monoclonal antibody secreted by the hybridoma, a double sandwich ELISA method for detecting tyrophagus putrescentiae is established, and basis is laid for diagnostic methods of accurately detecting Dermatophagoides farinae, such as tyrophagus putrescentiae, in houses, warehouses, fields and other environments.

The invention relates to the technical field of biology, and particularly discloses a preparation method and application of a dermatophagoides pteronyssinus allergen specific IgG Fab antibody fragment. The method comprises the following steps: 1) collecting and diluting serum, and filtering to obtain a serum sample; loading the sample on a protein A affinity chromatography column, and then eluting a serum IgG antibody to obtain a serum total IgG antibody; 2) adding dermatophagoides pteronyssinus to a affinity chromatographic column for coupling, adding the serum total IgG antibody in the step 1), and eluting the dermatophagoides pteronyssinus specific IgG antibody; and 3) adding agarose-linked papain into the house dust mite specific IgG antibody, uniformly mixing, incubating, centrifuging, and taking supernatant, thereby obtaining the house dust mite specific immunoglobulin G. The dermatophagoides pteronyssinus allergen specific IgG Fab antibody fragment can compete with IgE to bind to dermatophagoides pteronyssinus and reduce the formation of a dermatophagoides pteronyssinus-IgE compound, so that the binding of the dermatophagoides pteronyssinus-IgE compound to effector cells expressing IgE low-affinity receptors and IgE high-affinity receptors is reduced, the activation of sensitized cells is reduced, and the occurrence of inflammatory response is controlled.

The invention provides a method for inducing a mouse pruritus cutanea model by using dermatophagoides pteronyssinus protein and SEB, which comprises the following steps: adaptively feeding a mouse for 7 days, anesthetizing the mouse with isoflurane, shaving the back, smearing depilatory paste, scraping the depilatory paste and hair after 10 minutes, forming a 3-5cm < 2 > depilatory area, slightly scraping the depilated skin to cause skin lesion, and the like. According to the method, a mixed allergen is adopted to sensitize the local skin of the mouse, the total time of model preparation and model preliminary verification is 40 days, the model making period is shorter than that of 7 weeks of sensitization of a single allergen, and the model making severity is deeper. The mouse strain is common C57BL / 6 mouse, the price is low, and the model forming rate is 70% or above.

The invention belongs to the field of biotechnology, relating to a humanized monoclonal antibody IgG Fab fragment of dermatophagoides farinae 2 allergoid specificity. In the invention, by constructing a humanized immunoglobulin G gene library through a clone blotting method, an ELISA (Enzyme-Linked Immuno Sorbent Assay) sifts the humanized monoclonal antibody IgG Fab fragment of the dermatophagoides farinae 2 allergoid specificity from the library, through expression, purification and identification, the humanized monoclonal antibody IgG Fab fragment is proved to identify the dermatophagoidesfarinae 2 allergoid specificity and have higher affinity therewith and identify the specificity of dermatophagoides farinae crude antigen at the same time. A mast cell degranulation inhibition test shows that the fragment obviously inhibits the combination of dermatophagoides farinae allergen and the allergic disease patient serum IgE attached to the mast cell surface and plays the role of competitively inhibiting antigen combination. Early prevention can be carried out on allergic asthma patients through the Fab antibody fragment, and a medicament composition for preventing and treating the allergic asthma is further prepared.

The invention discloses a preparation method of a long-acting antibacterial anti-mite cotton fiber. The preparation method of the long-acting antibacterial anti-mite cotton fiber is characterized by comprising the following steps of (1) preparing absorbent cotton; (2) preparing a cotton fiber suspension; (3) carrying out binding reaction on organosilicon quaternary ammonium salt and the cotton fiber; (4) carrying out silver ion catalytic reaction; and (5) filtering and washing. The antibacterial and anti-mite cotton fiber prepared by the preparation method provided by the invention has the antibacterial rate of more than 99 percent for staphylococcus aureus, escherichia coli, candida albicans, klebsiella pneumoniae and the like, has the antibacterial rate of more than 99 percent for fungi such as trichophyton rubrum (dermatophytosis) and the like, has the repelling rate of more than 95 percent for dermatophagoides farinae according to the national standard GB / T 24253-2009 Evaluation of Textile Anti-mite Performance, and achieves a very strong anti-mite effect.

The invention belongs to biotechnics in particular a cultivation method of dust mite of room. The steps of said method comprises: collection dust with mites, little cultivation in early stage, a vast amount of cultivation in mast bottle by collecting mites with fine needle and repeat collection by saturated saline flotation method, to cultivate dust mites of room in controlled temperature and humidity, to reach effective performance. The invention avoids a lot amount of work and time, pollution when cultivating in open container, low production of mites, and the degree of purity is more than 90 % contrasted to low degree of purity by traditional method; and the cultivation base is easy to attained and in rational proportion.