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 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 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 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 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.

The invention relates to a real-time fluorescent PCR detection kit for bumblebee microsporidia and a detection method thereof, which are specially used for detecting bumblebee microsporidia. The kit comprises: (1) a positive standard; (2) negative control; (3) PCR reaction solution; (4) ExTaq enzyme; (5) sterile water. The invention has the following advantages: (1) good stability and specificity:detection of bumblebee microsporidia has high specificity, has no cross reaction with other microsporidia and bumblebee pathogens, and has good repeatability; (2) high sensitivity: the sensitivity can reach 20 copies/[mu] L; (3) easy and fast operation: the whole reaction can be completed within 2 hours. The kit can be used for detecting Nosema bombycis and differentiating and diagnosing the Nosema bombycis from other Nosema bombycis, and has important significance for early prevention and timely treatment of the disease. The kit can be used for detecting Nosema bombycis and differentiating and diagnosing the Nosema bombycis from other Nosema bombycis.

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 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 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.