146 results about "Vibrio cholera" patented technology

The present invention provides a gene chip for quickly detecting pathogens from food source, discloses the preparation method of said gene chip and provides 26 oligonucleotide probe sequences for detection. Said gene chip can quickly, accurately and high-effectively detect and identify the class of the pathogens in food, and its detection range includes staphylococcus aureus, Shiga's bacillus, salmonella, colibacillus 0157, bacillus proteus, mononuclear hyperplastic listerella, enterocolitis yersinia, aeruginous pseudomonads, vibrio parahaemolyticus, vibrio cholerae, bacillus cereus, beta hemolytic streptococcus, coconut fermentation pseudomonads, boticin, vibrio jejuni and bacillus perfringens, etc.

The invention relates to a complex enrichment medium used for salmonella, vibrio parahaemolyticus and comma bacillus and a method for preparing the same. The formulation components of the complex enrichment medium in weight portion are as follows: 0.5 to 1.5 portions of buffer peptone water, 1000 portions of distilled water, 1.0 to 5.0 portions of glucose, 1.0 to 5.0 portions of mannitol, 0.03 to 0.06 portion sodium pyruvate, 0.5 to 2.5 portions of anhydrous sodium sulfite, 1.0 to 10.0 portions of sodium citrate, 5.0 to 25.0 portions of sodium chloride, 1 to 5 portions of cholate, and 0.0005 to 0.0025 portion of potassium tellurite. The method comprises the following steps: firstly, adding the buffer peptone water and the like into the distilled water, and sterilizing the mixture after heating and melting; and secondly, cooling the mixture to 50 DEG C, and then adding the potassium tellurite into the mixture to be mixed evenly. The complex enrichment medium can integrate two steps of primary enrichment and selective enrichment of bacterial detection to shorten the enrichment time to 24 hours, can perform enrichment on three target pathogens at the same time, and can inhibit the growth of other microorganisms.

The invention relates to a gene chip and an application, which belongs to the biological detection field. The invention designs a set of detection probes by aiming at 11 types of common infectious diarrheal disease pathogen microorganism in clinic, and the gene chip containing the set of the probe. The detection probes comprises a vibrio parahaemolyticus probe, a vibrio vulnificus, a vibrio cholera probe, a vibrio alginolyticus probe, a vibrio furnissii probe, a shigella probe, an escherichia coli probe, an aeromonas probe, a salmonella probe, a campylobacteria probe, and a proteusbacillus vulgaris probe; the above mentioned probe sequence is shown as SEQ ID No: 1-117. The gene chip and probe can detect 11 types of common infectious diarrheal disease pathogen, the detection flux is high, the specialty is strong, the sensitivity is high, and the detection is rapid and effective.

The invention discloses a compound microbial agent which is prepared from nitrifying bacteria, nitrosomonas, single cell bacteria denitrification salt, salt denitrifying bacterium vibrio cholera, achromobacter denitrificans, bacillus thuringiensis and bacillus subtillis, wherein the total viable count is more than 5.9x10<10>cfu/g; the nitrifying bacteria and the nitrosomonas account for 10 to 20 percent of the total viable count; the single cell bacteria denitrification salt, the salt denitrifying bacterium vibrio cholera and achromobacter denitrificans account for 15 to 20 percent of the total viable count; the bacillus thuringiensis and the bacillus subtillis account for 5 to 10 percent of the total viable count. The compound microbial agent has the benefits that the compound microbial agent is fed at a time, and no bacteria is required to be kept added after the system is stabilized; therefore, the operation cost of sewage treatment is reduced, and the convenience of sewage treatment operation is enhanced.

The invention relates to a multiplex PCR-based synchronous and rapid method for detecting 13 pathogenic microorganisms in water, which comprises the steps of using multiplex PCR to simultaneously amplify gene-specific fragments of the 13 pathogenic microorganisms including escherichia coli, enterohaemorragic escherichia coli o157:h7, legionella pneumophila, salmonella enteritidis, shigella dysenteriae, staphyloccocus aureus, listeria monoeytogenes, helicobacter pylori, mycobacterium tuberculosis, klebsiella pneumonia, vibrio cholera, bacillus anthracis and yersinia pestis, and detecting PCR amplified products through agarose gel electrophoresis, thereby achieving synchronous and rapid detection for the 13 pathogenic microorganisms.

The invention provides a genetic chip and a detection kit related to major pathogenic microorganisms causing infectious diarrhea of human beings, which are mainly specific to 13 types of bacteria including lapactic bacillus coli/shigella, salmonella, comma bacillus, vibrio parahemolyticus, aeromona hydrophila, plesiomonas, virio hollisae, vibrio fluvialis, vibrio mimicus, Wauter's strains, vibriodamsela and vibrio furnissii. The genetic chip comprises a solid-phase carrier and an oligonucleotide probe fixed on the solid-phase carrier, wherein the oligonucleotide probe comprises DNA (Deoxyribonucleic Acid) fragments selected from genes including a gyrB gene, an ITS gene, a dnaJ gene, a toxR gene and the like with remarkable biological evolution advantages or complementary DNA fragments. By adopting the genetic chip and the detection kit provided by the invention, major pathogenic microorganisms causing infectious diarrhea of human beings can be detected. The genetic chip and the detection kit have the characteristics of easiness and convenience for operation, high flux, high accuracy, high repeatability and the like, and can be applied to clinical detection of inspection departments in hospitals.

The invention relates to a gene chip for detecting nine pathogenicity vibrios in marine products and a kit thereof. The gene chip comprises a solid phase carrier and an oligonucleotide probe, wherein the oligonucleotide probe includes one or more selected from the following nucleotide sequences: 1) DNA sequences selected from genes of a vibrio hollisae, a vibrio vulnificus, a vibrio cholera, a vibrio parahemolyticus, a vibrio harveyi, a vibrio alginolyticus, a vibrio furnissi, a vibrio mimicus and a vibrio damsel, 2) complementary DNA sequences of the DNA sequences selected in the DNA sequences in 1), 3) complementary RNA sequences of the selected DNA sequences in 1) or 2). The kit comprises the gene chips. The gene chip for detecting nine pathogenicity vibrios in marine products and the kit thereof provided by the invention are used for detecting the nine pathogenicity vibrios in marine products, and have the advantages of simplicity in operation, good sensitivity and strong repeatability.

The invention discloses a method for preparing questin by utilizing an ocean aspergillus flavipes HN4-13 bacterial strain. The method comprises the following steps: conducting separation and purification on antibacterial active substances generated through fermentation of the aspergillus flavipes HN4-13 bacterial strain according to silica-gel column chromatography, Sephadex LH-20 column chromatography, preparation of high performance liquid chromatography (PHPLC) and the like by taking vibrio harveyi as an indicator bacterium, so as to obtain a vibrio harveyi-preventing active compound HY2; indentifying the structure of the active compound HY2 according to spectra data such as ESI-MS, 1H-NMR and 13C-NMR, so as to determine that the active compound HY2 is questin. The questin is obtained through separation of a metabolic product of aspergillus flavipes HN4-13; experiments show that the questin can perform a certain inhibiting function of the pathogenic bacteria, such as vibrio harveyi, vibrio anguillarum, vibrio parahaemolyticus and vibrio cholerae, of aquatic products, and can be used for preparation of medicine preventing pathogenic vibrio of the aquatic products.

The invention relates to a loop-mediated isothermal amplification method for rapid detection of cholera toxin vibrio cholera. A reagent comprises a loop-mediated isothermal amplification reaction liquid, Bst DNA polymerase, and a chromogenic reagent, wherein the reaction liquid contains a reaction buffer liquid, dNTP, magnesium sulfate, an upstream inner primer 5-TGAATCCACGGCTCTTCCCT-TGGTTATGGATTGGCAGG-3, a downstream inner primer 5-GGTTGTGGGAATGCTCCAAG-ACTTTGGGTTTTTTCATCGC-3, an upstream outer primer 5- GATATTGCTCCAGCAGCA-3, a downstream outer primer 5-CGTCAAGGAATTTTACACCTAG-3, and betaine. The method for detecting the cholera toxin vibrio cholera comprises the steps of the extraction of bacterial DNA, the loop-mediated isothermal amplification of the cholera toxin vibrio cholera, and chromogenic detection. The method has the advantages of rapidness, strong specificity, high sensitivity, and low cost.

The invention discloses an LAMP kit for Vibrio cholera in an aquatic product. The kit comprises a reaction buffer solution, specific primers for detecting the Vibrio cholera, a BstDNA polymerase, a Vibrio cholera DNA positive template and ultrapure water; and the specific primers for detecting the Vibrio cholera comprise a pair of inner primers FIP and BIP and a pair of outer primers F3 and B3. The kit has the characteristics of strong specificity, high sensitivity, simplicity and fastness, and can be used for real-time rapid and accurate detection of Vibrio cholera of a base.

The invention discloses fluorescent quantitative polymerase chain reaction (PCR) primers and probes for detecting pathogenic bacteria of multiple aquatic products simultaneously and a detection method, and belongs to the field of molecular detection. Five groups of the fluorescent quantitative PCR primers and the probes are provided and can be used for detecting salmonella, listeria monocytogenes, vibrio parahaemolyticus, shigella flexneri and vibrio cholerae non-O1 respectively, wherein except the primers and the probes for listeria monocytogenes genes and the primers and the probes for vibrio parahaemolyticus genes cannot be used simultaneously, the pathogenic bacteria which can be detected by the corresponding primers and probes can be detected simultaneously by combining any other two groups or more than two groups of the primers and probes. The invention also discloses multiple fluorescent quantitative PCR amplification systems which are constructed by using the primers and the probes, so the primers and the probes can detect whether samples to be detected contain the corresponding pathogenic bacteria or not quickly and accurately, are convenient to operate and high in specificity, can be made into kits and the like and are applicable to the detection of the pathogenic bacteria of the subsidiary agricultural products, and a false positive result is avoided.

The invention relates to an immunomagnetic bead containing an OmpU antibody, preparation of the immunomagnetic bead and a method for capturing pathogenic vibrios and detecting the pathogenic vibrios through multiplex PCR by utilizing the immunomagnetic bead. PCR amplification of an OmpU gene is conducted by taking genomes of the vibrios as templates, pure OmpU recombinant protein is obtained to serve as an antigen to prepare the antibody, and the immunomagnetic bead containing the OmpU antibody is prepared through the coupling-washing process; the immunomagnetic bead can capture multiple vibrios, and then the vibrios are identified by multiplex PCR specific primers which are designed on the basis of a danJ gene. The immuomagnetic bead can simultaneously detect the vibrio parahaemolyticus, the vibrio cholera, the vibrio alginolyticus, the vibrio vulnificus and the vibrio mimicus at most and can be widely applied to environment and import and export food inspection. The immuomagnetic bead combines the specificity of immune separation and the high efficiency of PCR amplification, has the capacities of separation, concentration and enrichment, can efficiently separate and enrich the low-concentration pathogenic bacteria in a sample, reduces or eliminates inhibition and influences of the sample to PCR, improves detection limitation and has the very good application prospect.

The invention discloses a gene chip-based method for synchronously and rapidly detecting thirteen pathogenic microorganisms in a water body. The method is characterized in that specific gene segments of the pathogenic microorganisms are amplified simultaneously by adopting multiple polymerase chain reactions (PCR), and then, a detection result is obtained through gene chip hybridization and chip scanning image analysis. The detected thirteen pathogenic microorganisms comprise escherichia coli, Enterohemorrhagic Escherichia coli O157:H7, legionella pneumophila, salmonella enteritidis, Shigella, staphylococcus aureus, Listeria monocytogenes, Helicobacter Pylori, mycobacterium tuberculosis, Klebsiella pneumonia, vibrio cholera, Bacillus anthracis and Yersinia pestis.

The invention provides two primer pairs and further provides application of the two primer pairs for detecting or detecting vibrio parahaemolyticus and vibrio cholerae in an aided mode or preparing products for detecting or detecting vibrio parahaemolyticus and vibrio cholerae in an aided mode. The invention further provides a kit comprising the two primer pairs and application, and a method for detecting vibrio parahaemolyticus and vibrio cholerae on the basis of the two primer pairs or the kit. An experiment proves that the two primer pairs are good in specificity and high in sensitivity and detection efficiency, the established multi-DPO-PCR method for vibrio parahaemolyticus and vibrio cholerae on the basis of the two primer sets is high in throughput, sensitive, accurate and rapid, and the more effective method is provided for simultaneously detecting vibrio parahaemolyticus and vibrio cholera.

The invention relates to the technical field related to elimination of thalli, and particularly provides a preparation for strongly eliminating vibrios and a preparing method of the preparation. On one hand, provided is the preparation for strongly removing vibrios, wherein the preparation comprises the components in percentage by weight: 65-85% of a first guanidine derivative, 2-12% of a second guanidine derivative and the balance being ethanol, wherein the first guanidine derivative is polyhexamethylene guanidine salt and/or polyhexamethylene biguanide salt; and the relative molecular weightof the second guanidine derivative is 500-1500. The vibrios can be effectively removed by utilizing polyhexamethylene guanidine hydrochloride and chlorhexidine hydrochloride under the specific content. The preparation provided by the invention has a strong removal effect on pathogenic vibrios, such as vibrio parahaemolyticus, vibrio anguillarum, vibrio alginolyticus, vibrio cholerae, vibrio fluorescens and the like, has a good cracking effect on aeromonas hydrophila, aeromonas punctata, escherichia coli, salmonella and other gram-negative bacteria, and has a quite high removal rate after being used for 48 hours.

The invention belongs to the field of biological detection technology and relates to a GeXP analysis method for synchronous detection of vibrio cholera and vibrio parahaemolyticus in food. Primers used in the detection comprise a pair of vibrio cholera specific primers, a pair of vibrio parahaemolyticus specific primers and a pair of universal primers. The method comprises the following steps: carrying out enrichment culture on a sample to be detected, extracting nucleic acid, carrying out PCR amplification by the use of a specific primer pair and a fluorescently-labeled primer, detecting the amplification product by the use of a genetic analysis system, contrasting with standard molecular weight, and judging whether there is vibrio cholera and vibrio parahaemolyticus contamination in the detected sample according to detection atlas. Through verification, the detection system has characteristics of high specificity, good stability, short detection period, etc.