43 results about "Microsporidium species" patented technology

The invention discloses a group of primers and application thereof in quick detection of nosema bombycis. The primers have the nucleotide sequences as shown in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4. Based on the primers disclosed by the invention, related reagents applied to quick detection of nosema bombycis by an LAMP (Loop-Mediated Isothermal Amplification) method can be prepared by combining a conventional technology in the field, can detect the nosema bombycis by the LAMP method quickly and accurately, are suitable for operation on a small amount of sample and simple to operate, and have important significance in practical application.

The invention provides an important protein gene PTP1 relative with growth and infection of microsporidia of silkworms. The gene sequence CDS for coding the pole-tube protein PTP1 for microsporidia of silkworms is got based on the 9-magification gene data of microsporidia of silkworm and by collinear analysis. The pole-tube protein PTP1 got by the invention is of high importance for studying the breeding and infection mechanism of microsporidia, hence can play role in preventing and treating microsporidia in production practice. According RNAi interference, after the pole-tube protein PTP1 being interfered, the microsporidia loses the capability for infecting the host. The pole-tube protein PTP1 and its potential controlling gene of microsporidia will become a new factor for treating microsporidia infection. In addition, by means of transgenosis, the expression of the water-channel protein gene can be enriched, so that the water-channel protein gene can be used to control such destructive insects as locusts, etc.

The invention relates to a primer pair for rapidly detecting silkworm microsporidia and a reagent kit and a detection method thereof. The nucleotide sequence of the primer pair is shown as SEQ ID NO.7 and SEQ ID NO.8. By the utilization of the primer pair, different nucleic acid molecular markers are added into the upstream primer and the downstream primer respectively, PCR products of the marked primers can be directly determined through immuno-gold test strips, the operation steps are simplified, convenience is achieved, specificity is good, and sensitivity is high. Compared with a traditional gel electrophoresis method, the sensitiveness is increased by 10 times, and the primer pair can serve as an ideal detection means for silkworm microsporidia in silkworm finished product eggs and has good application prospects.

The invention discloses a direct PCR detection method of Bombyx mori nuclear polyhedrosis pathogeny BmNPV, comprising the following steps of: firstly, treating the blood of infected Bombyx mori by ethanol precipitation to remove a PCR reaction inhibitor; designing the PCR primer of a polyhedrin gene according to the genome analysis of the Bombyx mori nuclear polyhedrosis pathogeny BmNPV; carrying out PCR amplification on the polyhedrin gene, adding an anti-PCR reaction inhibitor BSA (Bull Serum Albumin) to a reaction system and simultaneously carrying out PCR reaction by the primer of a Bombyx mori chondriosome CO I gene to make positive control; respectively getting the blood of normal Bombyx mori and the inflected midguts of the Bombyx mori infected with white muscardine, green muscardine, a bacterial disease and a microsporidia disease and then carrying out PCR reaction specification analysis; and detecting the PCR reaction result by agar gel electrophoresis. The whole detection process has simple operation and can be completed only by 4 hours. The disease can be diagnosed at the early stage of BmNPV infection by adopting the direct PCR detection method to carry out periodical sampling detection so as to adopt a measure in time to prevent the disease from happening.

The invention discloses a multiplex PCR detection kit for newly-emerged intestinal protozoa, which is used for simultaneously detecting cryptosporidium, giardia intestinalis, cyclospora cayetanensis and microsporidia. The kit comprises the following primers: specific primers for cryptosporidium: shown in SEQ ID NO.1 and SEQ ID NO.2; specific primers for giardia intestinalis: shown in SEQ ID NO.3 and SEQ ID NO.4; specific primers for cyclospora cayetanensis: shown in SEQ ID NO.5 and SEQ ID NO.6; and specific primers for microsporidia: shown in SEQ ID NO.7 and SEQ ID NO.8. In addition, the invention also discloses a multiplex PCR detection method for cryptosporidium, giardia intestinalis, cyclospora cayetanensis and microsporidia. Various validations show that the multiplex PCR detection kit and detection method disclosed by the invention have the characteristics of rapidness, sensitivity, and strong specificity, and solve the cumbersome steps of multiplex PCR detection, and through one-time reaction, an effect of simultaneously detecting multiple genes and determining pathogens can be achieved, so that the workload is greatly reduced, the detection time is shortened, and the detection cost is reduced, therefore, the multiplex PCR detection kit and detection method disclosed by the invention have a greater development trend.

The invention discloses an ultra-low volume suspending agent containing nosema locustae and metarhizium anisopliae and used for grasshopper control as well as a preparation method of the suspending agent. The ultra-low volume suspending agent comprises raw materials in parts by weight as follows: 1-6 parts of metarhizium anisopliae spore powder, 1-10 parts of nosema locustae, 1-5 parts of a sulfonate anionic emulsifier, 0-5 parts of a non-ionic surfactant, 5-15 parts of vegetable oil, 50-90 parts of polyol and 0.001-0.1 parts of an ultraviolet protective agent. The ultra-low volume suspending agent combines nosema locustae and metarhizium anisopliae for use, so that respective defects of the nosema locustae and metarhizium anisopliae can be overcome, ideal control effect can be achieved, the flash point is high, volatilization is low, good liquidity is realized, requirements for ultra-low volume spray for the ground and an airplane can be met, phytotoxicity on plants is avoided, the environment is not polluted, the activity of spore and microsporidia can be better kept, the control effect is good, an aversion function is not produced, and antifeedant of pests is avoided.

The invention provides a light-stimulating chemiluminiscence immunological detection method of silkworm mature egg microsporidiosis. The light-stimulating chemiluminiscence immunological detection method comprises preparation of silkworm microsporidia sporoderm protein antigens, establishment of anti-rat silkworm microsporidia sporoderm protein monoclonal antibody hybridoma cell strains, ascites preparation and purification of silkworm microsporidia sporoderm protein monoclonal antibodies, and establishment of a silkworm microsporidia Alpha LISA detection method. When the method is used for detecting, monomer oxygen cannot be dispersed to receptor microspheres when biomolecules do not have specific mutual effect, and no signals are generated, so that the relatively high sensitivity and accuracy are realized, and a detection result is relatively accurate; the method has relatively high uniformity, less background interference and low background, and has a wide dynamic detection range; and when a kit is used for detecting, the demanded quantity of samples is extremely few and the samples do not need to be washed in a detection process, so that a detection flow is simplified and the market prospect is good.

The invention belongs to the field of breeding and particularly discloses a reagent box and a detection method. The reagent box comprises a 2x reacting, mixing and buffering solution, preferably designed upstream and downstream primers, a Taq DNA polymerase, a positive control solution, a negative control solution and ddH2O. The invention overcomes the defects of the method for detecting microsporidia of intestinal epithelial cells in the prior art and provides the novel reagent box and the detection method for rapidly detecting PCR (Polymerase Chain Reaction). The reagent box and the detection method are practicable and feasible in clinical diagnosis of microsporidia of intestinal epithelial cells of litopenaeus vannamei, have the advantages of high speed, accuracy and specificity, meet the requirements of clinical diagnosis and provide convenience for detecting the microsporidia of intestinal epithelial cells of litopenaeus vannamei.

The invention relates to a micro-microsporidia spore water suspension used for preventing and controlling locusts, and a preparation method thereof. The active component of the locust micro-microsporidia spore water suspension is locust micro-microsporidia spores, and proper amounts of Morwet D425 and xanthan gum are added. The locust micro-microsporidia spore water suspension has the advantages of simple preparation method, good leaf surface adhesiveness, high heat resistance, substantially improved dispersion and uniformity, normal temperature storage resistance, substantial prolongation of the shelf life, good locust killing effect, and cost reduction, is safe to chemical pesticides, and is important for improving the Chinese locust prevention and treatment technology level.

The invention discloses an Eriocheir sinensis Microsporidia in-situ hybridization detection probe and kit. The sequence of the probe is disclosed as SEQ ID NO.1. After the probe is labeled by digoxin and combined with Microsporidia 18S subunit ribosome DNA (18S SSU rDNA) by hybridization, the Microsporidia infection can be judged under a microscope after alkaline phosphatase detection system color development. The invention also discloses a kit containing the probe, which has favorable specific, is simple to operate and can intuitively combine the pathogen detection and pathological changes. On the premise of efficiently and accurately detecting the Spiroplasma, the kit can analyze the infection rate and infection intensity of the sample section. The hybridization between the probe and 18S SSU rDNA is specific, the signal is progressively amplified, and thus, the kit disclosed by the invention is more sensitive and accurate than the conventional dyeing observation detection. The detection result proves that the section subjected to hybridization color development can be stored for a long time.

The invention discloses a method for quickly quantifying the concentration of trehalose in a single microsporidia spore based on a single cell Raman spectrum, and belongs to the technical field of quantitative analysis. The method mainly adopts a single cell Raman spectrum technology and a pair of laser tweezers to quantitatively analyze the concentration of the trehalose in the single microsporidia spore. The method is easy and convenient to operate and pollution-free, and no chemical agents are needed; a spore sample is directly detected in germfree water, and the concentrations of the trehalose in the individual microsporidia spores from different sources can be quickly and intuitively quantified, so that heterogeneity of the concentration of the trehalose in the single spore in the same colony is obtained; the method has the advantages of simplicity, efficiency and reliability.

The invention discloses a method for quickly detecting microsporidia spores based on single cell Raman tweezers, belongs to the technical field of pathogenic bacterial detection. The method mainly adopts a pair of laser tweezers and a single cell Raman spectrum technology to judge spores of pathogenic bacteria microsporidia in a host body or the natural environment. The method is easy and convenient to operate, no staining is needed; detection is directly carried out in germfree water; furthermore, whether a sample contains the microsporidia spores or not can be quickly and intuitively judged; therefore, the method is worthy of being popularized and is simple, convenient, efficient and reliable for detection of the microsporidia spores.

The invention discloses a real-time fluorescent quantitative PCR primer pair for detecting hepatospora eriocheir. Sequences of the primer pair are shown as SEQ ID NO:1 and SEQ ID NO:2. The invention further discloses a real-time fluorescent quantitative PCR detection kit and application thereof. The primer pair, the kit and a detection method, provided by the invention, have the following advantages in detection of the hepatospora eriocheir: no cross-reaction occurs with other pathogenic microsporidia, and the specificity is strong; variation coefficients of repeatability test results within agroup and between groups are all less than 2%, and the repeatability is good; and the amplification efficiency is 98.3%, and the PCR primer pair can detect 2.87*101 copies / [mu]L at lowest, which is 100 times that of an ordinary PCR. The detection method established by the invention is higher in amplification efficiency and detection sensitivity; and a whole experimental process only takes 2 hoursfrom extraction of DNA from the to-be-detected hepatospora eriocheir to a result analysis, and the detection efficiency is greatly improved.

The present invention relates to Lymantria xylina cell lines established from pupal tissues of L. xylina Swinhoe, including NTU-LY-1, NTU-LY-2, NTU-LY-3, and NTU-LY-4. These four cell lines were confirmed to be newly established cell lines derived from L. xylina by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) and isozyme analysis. The genotypes and characteristics of the abovementioned cell lines are totally different from other insect cell lines. In addition, these four L. xylina cell lines are susceptible to insect baculovirus of L. xylina multiple nucleopolyhedrovirus (LyxyMNPV), LdMNPV-like virus, PenuMNPV, as well as microsporidia of Nosema sp. (isolated fromEurema blanda) and N. bombycis and the like. Therefore the invention can be applied in multiplication of the abovementioned insect-pathogenic microorganisms to produce biopesticides in pest control. In addition, the cell lines can also be used as hosts for the expression vectors of said baculovirus to produce recombinant proteins.

The invention discloses a method for early detection of grouper intestinal tract microsporidia and application thereof. The method includes the following steps that 1, a fore intestine of grouper is selected, and intestine content is scrapped; 2, genomic DNA of the fore intestine content of grouper is extracted; 3, with the extracted genomic DNA as a template, a specific primer is adopted to conduct PCR amplification; 4, a PCR amplification product in the step 3 is taken and subjected to electrophoretic analysis, a result is judged according to the size of the amplification product, if 450 bp stripes can be amplified through specific amplification, it is proved that the sample is infected by the grouper intestinal tract microsporidia. The problem that diagnosis is difficult as a grouper intestinal tract microsporidia individual is small is solved, programming and standardization of diagnosis are achieved, detection specificity and sensitivity are high, and the method can be applied to diagnosis of grouper intestinal tract microsporidia disease, applied to investigation of the molecular epidemiology and used for assessing whether aquaculture water and bait are suitable for grouper aquaculture or not.

A composition for controlling or preventing microsporidia in a fish or a seafood is prepared by using, as an active ingredient, at least one compound selected from the group consisting of compounds represented by the following chemical formula (I):(wherein, R2, R4, R5, R6 and R7 each independently represent a specific substituent), pharmaceutically acceptable salts thereof, and compounds generating the compounds represented by the chemical formula (I) by metabolism in the body of the fish or the seafood. By using this composition for controlling or preventing microsporidia in a fish to or a seafood, a method for controlling or preventing microsporidia in a fish or a seafood is provided, which prevents microsporidian infection in the muscle or organs of the fish or the seafood, and / or suppresses the growth of microsporidia in the muscle or organs of the fish or the seafood, and / or highly effective in eliminating microsporidia from the body of the fish or the seafood and also excellent in safety.

The present invention relates to Lymantria xylina cell lines established from pupal tissues of L. xylina Swinhoe, including NTU-LY-1, NTU-LY-2, NTU-LY-3, and NTU-LY-4. These four cell lines were confirmed to be newly established cell lines derived from L. xylina by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) and isozyme analysis. The genotypes and characteristics of the abovementioned cell lines are totally different from other insect cell lines. In addition, these four L. xylina cell lines are susceptible to insect baculovirus of L. xylina multiple nucleopolyhedrovirus (LyxyMNPV), LdMNPV-like virus, PenuMNPV, as well as microsporidia of Nosema sp. (isolated from Eurema blanda) and N. bombycis and the like. Therefore the invention can be applied in multiplication of the abovementioned insect-pathogenic microorganisms to produce biopesticides in pest control. In addition, the cell lines can also be used as hosts for the expression vectors of said baculovirus to produce recombinant proteins.

The invention discloses LAMP detection primers suitable for bombyx mori egg microsporidia and a quick detection method thereof. The detection primers are Septin1-F3, Septin1-B3, Septin1-FIP and Septin1-BIP, the four primers are represented as the SEQ ID No: 1-4. On the basis of the primers, detection steps and system are optimized, and the LAMP quick detection method for the bombyx mori egg microsporidia is disclosed. The method is simple in operation, is free of complex instruments and amplification program, is short in detection time and is low in interference on amplification due to impurities. A reaction result is easy to determine and has strong specificity. The invention provides important basic and technical basis for applying the detection on the bombyx mori egg microsporidia with the LAMP and ensuring health and toxic-free for silkworm eggs. The method can satisfy toxicity detection demands in academic institutions, silkworm egg producing departments and silkworm egg quality detection centers and is easy to promote in large scope.

The invention discloses a co-immunoprecipitation method suitable for nosema bombycis. The three technical problems in the prior art are solved, wherein as the first problem, spore total protein for a co-immunoprecipitation experiment is difficult to extract, and as the sporoderm of the nosema bombycis is thick, common protein extraction liquid cannot enter a spore, and protein cannot be fully extracted; as the second problem, due to the gravity, immune ball beads for co-immunoprecipitation can be naturally sunk to the bottom in the reacting process, stacked ball beads, protein liquid and antibodies cannot be fully combined, and the experiment result is influenced; as the third problem, most antibodies for the co-immunoprecipitation experiment are self-prepared polyclonal antibodies, the specific effect of the antibodies is poor, and nonspecific bands usually appear; in addition, detection to target protein can also be influenced by heavy chains and light chains of the antibodies. By means of the co-immunoprecipitation method suitable for the nosema bombycis, the specificity of the nosema-bombycis co-immunoprecipitation is improved, stability is high, and repeatability is good.

The invention discloses a macrobrachium rosenbergii microsporidia visualized quick detection kit and a detection method thereof. The macrobrachium rosenbergii microsporidia visualized quick detection kit comprises a microsporidia DNA extraction reagent and a reaction reagent, and the reaction reagent contains a nucleic acid primer set used for detecting macrobrachium rosenbergii microsporidia proliferation, wherein a primer group comprises a primer MrMSP-F3, a primer MrMSP-B3, a primer MrMSP-FIP, a primer MrMSP-BIP, a primer MrMSP-LpF and a primer MrMSP-LpB. According to the macrobrachium rosenbergii microsporidia visualized quick detection kit, a set of optimized quick proliferation reaction system is established, so that not only is macrobrachium rosenbergii microsporidia qualitative detection simpler and faster, high in specificity and high in sensitivity, but also the kit can be used for field detection, and has very high scientific study and economic value.

The invention discloses a group of microsporidium molecule universal detection primers and a kit thereof. The universal detection primers include an upstream primer HMG1F and a downstream primer HMG1R; the nucleotide sequence of the upstream primer is as shown in SEQ ID NO.3, and the nucleotide sequence of the downstream primer is as shown in SEQ ID NO.4. The detection primers target at the gene HMG1 and can be applied to detecting a plurality of microsporidia simultaneously, and the detection results are reliable, easy to operate and high in sensitivity; especially, the universal detection primers have extremely important practical application significance to early detection of various microsporidia in a sample in practical quarantine work.

The invention discloses a method for early detection of grouper intestinal tract microsporidia and application thereof. The method includes the following steps that 1, a fore intestine of grouper is selected, and intestine content is scrapped; 2, genomic DNA of the fore intestine content of grouper is extracted; 3, with the extracted genomic DNA as a template, a specific primer is adopted to conduct PCR amplification; 4, a PCR amplification product in the step 3 is taken and subjected to electrophoretic analysis, a result is judged according to the size of the amplification product, if 450 bp stripes can be amplified through specific amplification, it is proved that the sample is infected by the grouper intestinal tract microsporidia. The problem that diagnosis is difficult as a grouper intestinal tract microsporidia individual is small is solved, programming and standardization of diagnosis are achieved, detection specificity and sensitivity are high, and the method can be applied to diagnosis of grouper intestinal tract microsporidia disease, applied to investigation of the molecular epidemiology and used for assessing whether aquaculture water and bait are suitable for grouper aquaculture or not.

The invention discloses a pharmaceutical composition containing albendazole for resisting microsporidia for silkworms. The pharmaceutical composition is a medicine compound powder prepared from albendazole, enrofloxacin, a filler and an appropriate compound cosolvent. The pharmaceutical composition disclosed by the invention can be applied to prevention and treatment of a pebrine disease and bacteriosis, and comprises two application modes: directly spraying the medicine compound powder on mulberry leaves to feed silkworms, or blending the medicine compound powder with water, preparing a solution, soaking the mulberry leaves and airing to feed the silkworms. The administration time is continuous 12-15 days until mounting and cocooning of silkworm from the fourth larva of silkworm; and the composition is fed once a day except for a moulting stage. The application method disclosed by the invention is convenient, simple and relatively practical. A result proves that a pebrine disease of the silkworms can be effectively prevented and treated.

The present invention discloses a nosema bombycis DNA2 gene and application thereof. According to the invention, firstly cloning is carried out to acquire the nosema bombycis DNA2 gene with a full-length sequence shown as SEQ ID NO.1, an ORF rame sequence is shown as SEQ ID NO.2, and an amino acid sequence of encoding protein is shown as SEQ ID NO.3. On the basis, the invention also provides a specific detection primer set for specific detection of nosema bombycis, the specific detection primer set has very good specificity and sensitivity on nosema bombycis, and has very good application prospect in specific detection of nosema bombycis. Moreover, as a newly discovered protein gene, the nosema bombycis DNA2 gene has potential functional application. Discovery of the DNA2 gene provided by the invention has certain significance for theoretical verification and production practice of treatment and detection.

The invention relates to a Chinese herbal compound preparation used to prevent and control water chest disease in river crab breeding, and belongs to the technical field of aquaculture. It mainly takes honeysuckle, gallnut, and artemisia annua, weighs them in proportion and then adds water to decoct them. The decoction liquid is mixed evenly with salicylic acid, citric acid and polyhexamethylene biguanide in proportion by weight, divided into packages, and finally packaged and stored in the warehouse. The present invention uses Chinese herbal plants that are safe and non-toxic to river crabs, and is combined with various substances such as organic acids, polymer environmentally friendly fungicides, etc. It has antibacterial and antiviral properties, kills microsporidians, eliminates toxins in the breeding environment, and prevents and promotes disease. It is grown in one body, has clear ingredients, and has a wide range of effects. It can be poured directly, is easy to use, has high bioavailability, and is not affected by environmental conditions. After use, it can quickly repair the ecological environment of river crab breeding and reduce the residues of heavy metals and pesticides in the breeding environment. It can effectively reduce the number of microsporidians, bacteria and viruses in the breeding environment, reduce the incidence of "water mites" disease in river crabs, protect body tissues, promote growth, and improve the breeding output and quality of river crabs.

The invention discloses a silkworm microsporidia HMG1 gene as well as a specific primer set used for rapidly detecting silkworm microsporidia and application thereof. The primer set comprises an upstream primer HMG1-s and a downstream primer HMG1-sR, wherein a nucleotide sequence of the upstream primer is shown in SEQ ID No.5, and a nucleotide sequence of the lower primer is shown in SEQ ID NO.6. A target gene of the detection primer is a high mobility protein gene HMG1 related to sex of the silkworm microsporidia, when the high mobility protein gene HMG1 is taken as the target gene for a silkworm tissue, especially silkworm egg microsporidia molecular detection, the characteristics of reliable detection result, easy operation, strong specificity and high sensitivity can be realized, and the high mobility protein gene HMG1 can be applied to high-sensitivity and rapid silkworm microsporidia molecular detection, especially early detection of the silkworm microsporidia, and has great significance in practical application.

The invention provides a water-channel protein gene relative with sprouting of microsporidia of silkworms. The gene sequence CDS for coding the water-channel protein NbAQP for microsporidia of silkworms is retrieved based on the 9-magification gene data of microsporidia of silkworm, and is proved by a series of biologic cloning technologies. The water-channel protein NbAQP got by the invention is of high importance for studying the sprouting mechanism of microsporidia, hence can play role in preventing and treating microsporidia in production practice. According RNAi interference, after the water-channel protein NbAQP being interfered, the microsporidia loses the capability for infecting the host, this provides a forceful means for preventing and treating granulosis of silkworms; in addition, by means of transgenosis, the expression of the water-channel protein gene can be enriched, so that the water-channel protein gene can be used to control such destructive insects as locusts, etc.

The invention relates to a method for detecting microsporidiosis of locust by PCR (Polymerase Chain Reaction), and provides PCR primers and a kit for detecting microsporidiosis of locust. The method for detecting the microsporidiosis of locust by PCR is capable of performing PCR detection on locust by utilizing the primers or the kit. By means of the content disclosed by the invention, the situation of locust infected by microsporidia can be quickly determined on a large scale, and the number of samples required by detection is greatly reduced, the detection accuracy is greatly improved, the results are in the credibility, and the sensitivity can be 7*10<3> spore / ml. Compared with a traditional microscopy, the method disclosed by the invention has the advantage that the detection cost and workload can be greatly reduced.

The invention relates to a primer pair for rapidly detecting silkworm microsporidia and a reagent kit and a detection method thereof. The nucleotide sequence of the primer pair is shown as SEQ ID NO.7 and SEQ ID NO.8. By the utilization of the primer pair, different nucleic acid molecular markers are added into the upstream primer and the downstream primer respectively, PCR products of the marked primers can be directly determined through immuno-gold test strips, the operation steps are simplified, convenience is achieved, specificity is good, and sensitivity is high. Compared with a traditional gel electrophoresis method, the sensitiveness is increased by 10 times, and the primer pair can serve as an ideal detection means for silkworm microsporidia in silkworm finished product eggs and has good application prospects.