18 results about "Microsporidium" patented technology

Provided are chitosan-RNA particles comprising low molecular weight chitosan and ssRNA or dsRNA, as well as compositions and cultured aquatic animals comprising the chitosan-RNA particles, and methods of use thereof in the treatment or prevention of microsporidia infection in aquaculture, the ssRNA or dsRNA is partially complementary, bound, or at least 90% identical to the mRNA target of a pathogenic microsporidium parasite in cultured fish or crustacean.

The invention discloses a method for breeding, separating and purifying nosema bombycis, which comprises the following steps: infecting 2-year-old silkworms for 6-12 hours by using nosema bombycis with the concentration of 1.0 * 10 < 5 > spores / mL to 5.0 * 10 < 5 > spores / mL, feeding the silkworms with clean mulberry leaves until the silkworms fall asleep at 4 years old, and gradually causing pebrine death when the silkworms grow at 5 years old; the method comprises the following steps: grinding dead silkworms to prepare grinding liquid, filtering the grinding liquid, and separating out filtrate enriched with nosema bombycis; centrifuging the filtrate to obtain a precipitate which comprises grey silkworm body tissues on the upper layer and white nosema bombycis on the lower layer, shaking the centrifuge tube by using a shaking table to release the grey silkworm body tissues to supernate, pouring out the supernate, and retaining the white nosema bombycis at the bottom of the centrifuge tube. According to the method, the nosema bombycis is bred, separated and purified according to the growth rules and characteristics of the bombyx mori and the nosema bombycis, a high-quality sample can be efficiently obtained, and the nosema bombycis is high in content and few in impurity.

The invention discloses a locust microsporidia strain and a preparation and use method thereof, and relates to the technical field of locusts, the locust microsporidia strain and locust microsporidia have the preservation number of CGMCC No.19390 and are named as PL-GM1 strain. The preparation method of the locust microsporidia strain comprises the following steps: S1, screening high-temperature-resistant mutant strains; s2, other high-temperature-resistant microorganisms are used in a combined mode; s3, cultivating healthy insects; s4, healthy insect inoculation; s5, feeding diseases and pests; s6, microspores are processed and inspected, packaging and quality inspection are conducted, and finally freezing storage is conducted. According to the method, high-temperature-resistant mutant strains are screened, high-temperature-resistant culture can be conducted on the locust microsporidia of the PL-GM1 strain, and the locust microsporidia of the PL-GM1 strain can have the high-temperature-resistant characteristic; other high-temperature-resistant microorganisms are used in a combined mode, the microsporidia screened in the step S1 and the other high-temperature-resistant microorganisms are co-cultured, and the high-temperature-resistant capacity and the prevention and treatment effect of the microsporidia are enhanced.

The invention relates to the technical field of monoclonal antibodies, in particular to a monoclonal antibody targeting EcSSP1 protein and application of the monoclonal antibody in detection of microsporidia. The amino acid sequence of a heavy chain variable region of the monoclonal antibody provided by the invention is as shown in SEQ ID NO.9, and the amino acid sequence of a light chain variable region of the monoclonal antibody is as shown in SEQ ID NO.10. The monoclonal antibody provided by the invention is obtained by screening by taking the recombinant protein EcSSP1 as an immunogen, has relatively high specificity, can be positioned on the surface of microsporidia, has relatively high detection accuracy and sensitivity, can be used for detecting in-vivo microsporidia in the early stage of infection, and can be used for detecting microsporidia in vivo. And if the antibody is positive, the current infection of the microsporidia can be confirmed, the operation is simple and convenient, the subcellular localization research of the microsporidia protein can be carried out by utilizing a multicolor fluorescence labeling method, and the application prospect is relatively good.

The invention discloses application of a bombyx mori key gene circ0000157, and belongs to the field of biotechnology research. By analyzing the biogenesis mechanism of the circ0000157, it is proved that the circ0000157 is formed by reverse shearing of an exon and an intron. The circularity of circRNA is verified through agarose gel electrophoresis, Sanger sequencing and exonuclease digestion, it is verified through experiments that proliferation of the nosema bombycis can be inhibited by inhibiting expression of the key gene circ0000157 of the nosema bombycis in a host, and it is proved that the circ0000157 serves as a molecular sponge to adjust the miR-281-5p to affect proliferation of the nosema bombycis. A foundation is laid for analysis of an infection mechanism of the nosema bombycis, and an important target is provided for creation of resistance materials.

The invention provides application of vitamin B2 in preparation of a product for inhibiting or killing microsporidia, and belongs to the technical field of cultivation. The invention provides application of vitamin B2 (VB2) in preparation of a product for inhibiting or killing microsporidia. The VB2 can enable mature spores of the microsporidia to lose activity, so that the infection of the microsporidia is inhibited. From the perspective of safety, a proper amount of VB2 is harmless to human and animals, and can be used as a nutritional additive for animals. Moreover, the VB2 is used as a water-soluble vitamin necessary for organisms, the metabolic pathway of the VB2 is thoroughly researched, and the VB2 can participate in an oxidation-reduction reaction through a flavoenzyme system (such as FAD / FMN) and is naturally discharged out of a body without a residual risk. In addition, the VB2 keeps broad spectrum when inhibiting microsporidia, and is non-toxic to bred animals at the same time.

This invention relates to a triple nano-PCR detection method for silkworm microsporidia, Beauveria bassiana, and nucleopolyhedrovirus, belonging to the field of molecular diagnostic technology. Addressing the technical problem of existing silkworm disease diagnostic methods' inability to simultaneously, efficiently, and sensitively identify these three commonly mixed infection pathogens, this invention provides an integrated solution. The key technical points include extracting DNA from the sample to be tested, using this DNA as a template, performing nano-PCR amplification in the same reaction system using a specific primer set, and subsequently detecting the amplification products by agarose gel electrophoresis. The infection status of silkworm microsporidia, Beauveria bassiana, and nucleopolyhedrovirus is determined based on the specific bands at 733 bp, 230 bp, and 860 bp, respectively. This method is mainly used for the rapid, simultaneous diagnosis and identification of mixed infections of microsporidia, Beauveria bassiana, and hemorrhagic septicemia during silkworm rearing.

The present invention relates to a novel simultaneous multiple all-in-one LOP molecular diagnostic platform structure and a simultaneous multiple molecular diagnostic kit for diagnosing shrimp white spot virus, acute hepatopancreatic necrosis, and microspore infection using same. According to the present invention, shrimp white spot virus, acute hepatopancreatic necrosis, and microspore infection that necessitated PCR analysis can now be easily and simultaneously detected on-site via simultaneous multiple detection.

The present invention relates to the technical field of Microsporidia cryopreservation. Provided is a cryoprotectant for Enterocytozoon hepatopenaei. The cryoprotectant is mainly prepared from glycerol, dimethyl sulfoxide, and propylene glycol, with PBS buffer as a base solution. A volume percentage ratio of the raw materials dimethyl sulfoxide, glycerol, propylene glycol and PBS buffer is 10%:10%:10%:70%. The raw materials of the cryoprotectant are preferred, such that the obtained cryoprotectant achieves the optimal effect in indicators such as the mortality rate of Enterocytozoon hepatopenaei, the infection load in healthy shrimps, and the influence on polar tube protein 2 (EHP-ptp2) and heat shock protein 70 (hsp70). The present invention lays the foundation for achieving indoor preservation of Enterocytozoon hepatopenaei. The present invention solves the technical problem in the prior art that no cryoprotectant or preservation technique suitable for Enterocytozoon hepatopenaei is currently available.

The application is designed and screened according to the specific amplification primer of the conserved sequence of Chinese mitten crab microsporidium, the specific amplification primer screened is modified with a marker, and then the MIRA reaction condition is optimized; the MIRA detection technology has obvious advantages such as no need for thermal cycling, short detection time, simple operation, high sensitivity, simple equipment configuration and the like; compared with ordinary PCR detection, the MIRA detection technology of Chinese mitten crab microsporidium established in the experiment can complete the reaction at 37 DEG C in only 25 min, the minimum detection limit is 1 copies / µL, which is 10 4 copies / µL higher than that of ordinary PCR, the detection accuracy reaches 100%, and good specificity is achieved.

The invention discloses a rabbit encephalitis microsporidia nested PCR specific primer as well as a detection method and application thereof, and relates to the technical field of rabbit encephalitis microsporidia detection. The invention relates to a nested PCR (Polymerase Chain Reaction) specific primer for rabbit encephalitis microsporidia. A gene sequence of an outer forward outer primer is shown as SEQ ID NO.2; the gene sequence of the outer reverse outer primer is as shown in SEQ ID NO.3; the gene sequence of the inner side forward inner primer is as shown in SEQ ID NO. 4; and the gene sequence of the inner side forward outer and inner primers is as shown in SEQ ID NO. 5. The nested PCR primer designed by the invention is high in specificity, can only identify E.cloniculi, has no cross reaction with related species, has sensitivity improved by 100 times compared with that of conventional PCR, can detect more low-concentration positive samples, and reduces the omission ratio; the matched kit fills the market blank, and provides efficient and accurate technical support for clinic and prevention and control.

The invention discloses a fluorescent RAA primer group, a kit, application and a method for detecting common pathogen beauveria bassiana of bombyx mori, the kit comprises a specific primer group Bb-F / Bb-R, a probe Bb-P and a targeted beauveria bassiana ITS conserved region, during operation, only sample DNA needs to be extracted, reaction is performed for 30 minutes under the constant temperature condition of 42 DEG C, and a result is interpreted in real time through a fluorescent signal. The kit is high in specificity, only amplifies beauveria bassiana DNA, has no cross reaction on common silkworm pathogens such as silkworm nuclear polyhedrosis virus and silkworm microsporidia and healthy silkworm tissues, and also has no cross reaction on other fungal pathogens of silkworms; the sensitivity is high, and the lowest detection line is 3.91 * 10 copies / mu L; the method has the advantages of simple operation, fast detection, completion within 30 minutes, no need of complex equipment, suitableness for the fields of silkworm white muscardine silkworm early detection, epidemic situation monitoring, silkworm larva medicinal material inspection and the like, provides an efficient and accurate molecular diagnosis tool for silkworm white muscardine silkworm prevention and control and silkworm larva identification, and is especially suitable for being popularized and used in grassroots.

The invention provides a method, a kit and a system for detecting intestinal microsporidia based on a CRISPR-Cas12a technology, belongs to the field of nucleic acid detection, and aims to solve the problems that an existing intestinal microsporidia detection method is time-consuming, high in cost and low in sensitivity and specificity, and an existing CRISPR technology is poor in adaptability, insufficient in detection of low DNA template quantity and the like. Recombinase polymerase amplification (RPA) and nested polymerase chain reaction (PCR) are combined, a conserved genome sequence of the enterosporidia is taken as a target, high sensitivity of nucleic acid amplification and high specificity and signal amplification capability of CRISPR-Cas12a are fused, the lowest detection limit reaches 1021M, the lowest detection limit is far better than the single use effect of traditional PCR or CRISPR-Cas12a, ultra-sensitive, high-specificity, rapid and low-cost detection is realized, and the kit is suitable for clinical application. The kit is suitable for detecting enterosporidium pichia early infection or samples with low DNA template quantity.

This document discloses methods and compositions for use in various therapeutic, preventive, and control measures comprising from about 0.00001% by weight to about 100% by weight of an extract derived from sugarcane. The therapeutic, preventive, and control measures involve administering the extract to a subject, the extract comprising from about 10 catechin equivalents (CE) g / L to about 50 CE g / L of polyphenols or from about 100 CE mg / g to about 500 CE mg / g of polyphenols. These therapeutic and preventive measures include antimicrobial activity, particularly against respiratory viruses in humans, and treatment of microsporidia infections, such as EHP in shrimp and prawns. Control measures include reducing methane production in ruminants. The use of these compositions in, for example, dairy cows improves cow health by minimizing mastitis while increasing milk yield.

The invention discloses a method for monitoring microsporidia infection in a silkworm egg production process, which specifically comprises the following steps: collecting silkworm excrement or silkworm eggs as a to-be-detected object, adding saturated salt water into the to-be-detected object, grinding, filtering, standing filtrate, taking supernatant, observing, and if ellipsoidal spores which can swing up and down exist in the supernatant, judging that microsporidia infection is caused. If yes, the microsporidia pathogens exist in the excrement or the silkworm eggs. The invention further provides a kit which is suitable for DNA detection and verification of various common insect microsporidia in the mulberry field, including silkworm microsporidia. According to the invention, it is found for the first time that spores of nosema bombycis float in saturated salt water and swing obviously, the spores of nosema bombycis in silkworm excrement or silkworm eggs are enriched by using the saturated salt water, and whether the spores swing or not is observed under an optical microscope, so that whether the silkworm excrement or silkworm eggs contain microspore pathogens or not can be judged. And then the infection and the infection degree of the microsporidia can be determined through specific amplification of nucleic acid of the microsporidia.

Particulate compositions comprising magnesium and low molecular weight chitosan and ss or dsRNA partially complementary to, binding to or at least 90% identical to mRNA targets of microsporidia and viruses pathogenic in farmed crustaceans, compositions and farmed aquatic crustaceans comprising the same, and methods for their use in treating or preventing parasitic microsporidia and viral infection in aquaculture are provided.

The invention relates to an epitope polypeptide of cryptosporidium parvum Gp900 protein and application of the epitope polypeptide, the amino acid sequence of the epitope polypeptide is shown as SEQ ID NO.1, and the nucleotide sequence for coding the epitope polypeptide is shown as SEQ ID NO.2. Experiments prove that the epitope polypeptide has good in-vitro affinity with mouse H-2Db and has certain stability; after the epitope polypeptide is emulsified by an adjuvant to immunize a mouse, not only can CD8 + T cell immune response of the mouse be obviously stimulated, but also polypeptide specific CD4 + T cell response can be induced to be generated; compared with negative control, P is less than 0.01, and the IFN-gamma and the TNF-alpha generated by CD8 + T cells and CD4 + T cells induced by the Gp900 protein have extremely significant difference, so that the Gp900 protein and the related polypeptides thereof have corresponding biological immunology functions, have huge development and application potential in the field of cryptosporidium parvum specific immunotherapy, and have a broad application prospect in the field of cryptosporidium parvum specific immunotherapy. And a new direction is provided for preparing cryptosporidium parvum vaccines based on T cell epitopes.

The invention discloses an application of a micromolecule Z1444019930 of targeted nosema bombycis aquaporin in preparation of a medicine for preventing and treating nosema bombycis, and belongs to the field of molecular biology and pathology. When the use final concentration of Z1444019930 in cells is more than 1 mu M, the Z1444019930 can inhibit the germination of nosema bombycis, and when the Z1444019930 is added to bombyx mori with the concentration of more than 1 mM along with mulberry leaves, the morbidity of the nosema bombycis can be obviously reduced. The method can be used for research, development and production of medicines for preventing and treating pebrine of silkworms, and has application and popularization values.