43 results about "Enrichment broth" patented technology

The invention relates to a method for separating and purifying sulfate-reducing bacteria in oilfield production water: adding oilfield production water into a liquid culture medium for sulfate-reducing bacteria, and carrying out anaerobic enrichment culture at a constant temperature of 30-37°C; enriching culture solution for sulfate-reducing bacteria Carry out gradient dilution and solid interlayer plate coating, pour the culture medium of solid sulfate-reducing bacteria into the lid of the culture dish, cool and solidify to form the lower layer of culture medium, absorb the diluted enriched bacteria solution and spread it evenly on the lower layer for cultivation Then pour the upper layer of solid sulfate-reducing bacteria culture medium, and cover the bottom of the petri dish on the upper layer of culture medium for constant temperature cultivation; when a single colony appears to turn the medium black, pick the single colony into sulfate-reducing bacteria The liquid enrichment medium is used for enrichment culture, and the strain whose culture medium becomes the darkest is taken and the steps of sandwich plate to liquid enrichment culture are repeated three times to obtain pure strains of sulfate-reducing bacteria. The invention has the advantages of convenient operation, readily available materials, good separation and purification effects and the like.

The invention relates to an application of a Pseudomonas aeruginosa polyclonal antibody during a salmonella separation process. Acceding to the invention, the polyclonal antibody is used for detecting salmonella, the method comprises the following steps: preparing the polyclonal antibody of interference bacteria-Pseudomonas aeruginosa during the salmonella separation process, purifying by a saturation ammonium sulfate method, adding the purified antibody in a selective enrichment broth during the salmonella separation process, and reacting for 12 hours at the temperature of 37 DEG C and then carrying out subsequent separating culture.

The invention aims at a kit for shigella in enteric pathogenic bacterias as well as a use method for the kit, and mainly solves technical problems of high omission factor, complex identifying steps, higher cost and insufficiently visualized screened results in the existing method for separating the shigella. The kit comprises a universal type nonselective enrichment broth, a 10-time concentrated universal type nonselective enrichment broth, a shigella enrichment broth, a xylose lysine deoxycholate agar plate, an enzyme developing agar plate, a shigella biochemical identification tube, a hydrogen sulfide filter paper strip and an indole filter paper strip, and is characterized by further comprising liquid paraffin oil. All shigellas are identified through a reaction combination test of a substrate and a typical colonial morphologic, simple biochemical and specific enzyme correspondingly grown in a culture medium for selective separation.

The invention relates to a method for separating and identifying yersinia enterocolitica belonging to yersinia in enteric pathogenic bacteria, mainly solving the technical difficult problems of high missed detection rate, complex identifying steps, higher cost and insufficiently visualized screened results in the existing method for separating and identifying the yersinia enterocolitica. A kit comprises 1) general type nonselective enrichment broth, 2) 10-time concentrated general type nonselective enrichment broth, 3) yersinia cold enrichment broth, 4) yersinia quick selective enrichment broth, 5) a selective separation plate, 6) an enterotoxigenic yersinia enterocolitica developing agar plate, 7) a yersinia enterocolitica biochemical identification tube formula, 8) a hydrogen sulfide filter paper strip, and 9) an indole filter paper strip. The yersinia enterocolitica is separated and identified by using combination test of typical colonial morphologogy correspondingly growing by using selective enrichment and selective isolation culture medium, and simple biochemistry and specific enzyme-substrate reaction.

The solid-phase pouring enrichment method is to replace the liquid enrichment solution with solid-phase enrichment medium and swabs containing solid-phase enrichment medium, and to cultivate and isolate pathogenic bacteria from the original sample containing miscellaneous bacteria by pouring culture method . The supporting device is a container containing solid-phase enrichment medium and a swab containing medium, so that it has the functions of sampling, pre-enrichment, enrichment, and biochemical rough screening at the same time. Take the Salmonella detection tube used in the physical examination of practitioners as an example, moisten the anal swab containing the culture medium with water, directly sample and culture, and directly observe the reaction. If Salmonella is suspected, there will be colored suspicious spots on the swab. Take the spots for pure isolation, and then do further identification. Those without spots are negative, and there is no need to transfer to the plate. The detection rate of this method is significantly higher than that of the conventional method, and because the observation time can be extended, the delayed growth bacteria will not be missed. This method simplifies procedures, reduces costs, and has a high detection rate. Another Salmonella detection bag according to the same principle has a higher detection rate. A series of multi-functional products can also be developed, such as bacteria monitoring in food, food poisoning detection, infectious disease detection, nosocomial infection monitoring and other products.

The invention discloses a method for detecting staphylococcus aureus in food. The method comprises the following steps: preparing a culture medium, wherein the culture medium comprises a yeast extract, soytone, agar, sodium chloride, sodium pyruvate, glycine, lithium chloride, 4-methylum-belliferyl-beta-D-glucoside, p-nitro beta-D-glucoside and paranitrophenol-alpha-D-galactopyranose; (2) putting 10 g of a sample to be detected into 100 ml of the culture medium, and culturing for 36 hours at 37 DEG C so as to obtain enrichment broth; when naked eye observation shows that the enrichment broth is a yellow bacterial colony color, sucking the enrichment broth into a sterile injector, filtering by using a filtering membrane, recycling the filtrate, replacing the sterile injector, washing for 1-3 times by using PBS, washing for 1-3 times by using PBST, and observing whether the washing has fluorescent light under ultraviolet light or not, wherein the sample has staphylococcus aureus if the fluorescent light is observed. The method is rapid, simple and accurate.

The invention provides a culture medium capable of improving biological stuffing microbial activity and a preparation method thereof, and relates to the technical field of biological stuffing. The culture medium is characterized by being prepared from, by weight, 100-150 parts of peptone, 80-100 parts of beef extract, 10-20 parts of yeast extract, 20-25 parts of glucose, 5-10 parts of sodium chloride, 20-30 parts of agar, 15-20 parts of bile salt from a pig, 12-18 parts of casein hydrolysate, 7-12 parts of sodium nitrate, 5-8 parts of monopotassium phosphate, 10-12 parts of amino acid and 16-20 parts of enrichment broth. The prepared culture medium allows growing microorganisms to be rich in type, a proper survival environment is provided for the microorganisms, nutritional ingredients (N, P, K and trace elements) are reasonable, the nutritional ingredients needed in the microorganism growth and reproduction processes are met, the microbial activity is greatly improved, the odor treatment effect is good, and the strictest environmental requirement in various regions can be met in any season.

An offshore escherichia coli detection method is provided. According to the method, escherichia coli in seawater is intercepted onto a filter membrane by a suction filtration system, the filter membrane is put into a culture medium and cultured and is a microfiltration membrane having a pore diameter of 0.45 [mu]m, a bile lactose enrichment broth is added into a detection solution to perform bacteria-proliferating operation for 30-48 h, and if the solution is muddy and a gas is generated, it is determined that the escherichia coli is detected. The culture medium is an eosin-methylene blue agar plate, the culture time is 10-12 d, and colony growth conditions of the escherichia coli are observed after culturing. If a typical escherichia coli colony grows and is separated from the eosin-methylene blue agar plate, it is determined that the escherichia coli are detected. Compared with traditional microbe detection methods, the method is low in cost, rapid, convenient, simple, and high in sensitivity.

The present invention discloses a simple method for screening aquatic source aeromonas. The method includes that: a sample to be detected is subjected to enrichment culture in salt free alkali peptone water, wherein the culture condition is a conventional culture condition of the aquatic source aeromonas; enrichment broth is inoculated to an RS medium flat plate for culturing, wherein the culture condition is the conventional culture condition of the aquatic source aeromonas, if yellow and smooth bacterial colonies appear on the RS medium flat plate, the sample is suspected bacteria of aeromonas; the suspected bacteria of aeromonas is inoculated to a tryptone soybean agar medium flat plate for culturing, wherein the culture condition is the conventional culture condition of the aquatic source aeromonas, oxidase activity of the cultured bacterial colony is detected; if the bacterial colony can grow well in the salt free alkali peptone water, the bacterial colony appears yellow and smooth on the RS medium flat plate, and the oxidase test is positive, then the bacterial colony is determined to be the aquatic source aeromonas, and the sample to be detected contains the aquatic source aeromonas.

The invention relates to a quick extraction kit for common pathogenic bacterium genomes in food and a use method. The kit comprises carboxyl-modified silica-coated magnetic beads, a pretreatment buffer solution, a pyrolysis buffer solution, a rinsing solution, proteinase K, nucleic acid dissolution solution and the like. The use method of the quick extraction kit for common pathogenic bacterium genomes in food comprises steps as follows: a to-be-extracted sample is washed with the pretreatment buffer solution for removal of food substrate, enrichment broth and other impurities, the pyrolysis solution, proteinase K and the carboxyl-modified silica-coated magnetic beads are added to pyrolyse cells and release nucleic acid, the carboxyl-modified silica-coated magnetic beads are adsorbed on a magnetic grate after isopropyl alcohol is added, and high-purity nucleic acid is obtained through further purification. When the kit is used for extracting the common pathogenic bacterium genomes in food, the sample is not required to be pyrolyzed in advance, the effect of food substrate is smaller, extraction is quick, and the time and the cost are saved for laboratory detection.

Methods and kits for detection of bacteria, especially Vibrio parahaemolyticus and Vibrio vulnificus, are provided using a unique combination of selective ingredients and two-phase culture (solid-phase culture gel and liquid-phase culture/enrichment broth) allows for high sensitivity and specificity of the kits for growth of Vibrio parahaemolyticus and Vibrio vulnificus in the detection methods and kits. The invention, through the detection mechanism accomplished by the novel formulation of selective ingredients and the two-phase culture, allows for real-time detection of a single cell of Vibrio parahaemolyticus and Vibrio vulnificus within 24±2 hours of introducing a target sample to the Vibrio parahaemolyticus and Vibrio vulnificus detection kits.

The invention relates to a detection kit for Campylobacter jejuni in pork. The detection kit comprises a culture medium and a detection solution, wherein the culture medium is broth. The detection solution is composed of 3-5 mu l of 10 PCR (polymerase chain reaction) buffer solution, 0.2-0.4 mu l of Taq DNA (deoxyribonucleic acid) polymerase, 3-5 mu l of dNTP (deoxyribonucleotide triphosphate), 0.1-0.6 mu l of 100bp DNA Ladder Marker, 1-2 mu l of PCR primer, 5-10 ml of Tris.cl, 1-2.2mg of sodium chloride, 1-2.2mg of magnesium chloride, 0.2-0.5mg of disodium hydrogen phosphate dodecahydrate and 3-5mg of enrichment broth. The detection kit is an improvement on the basis of the conventional PCR technology, and can be used for qualitative detection of Campylobacter jejuni. By integrating the advantages of PCR, DHPLC (denaturing high performance liquid chromatography) and other techniques, the detection kit has the advantages of high specificity of detection results and high sensitivity, and ensures the effective detection of Campylobacter jejuni in food.

The invention relates to a kit for separating and identifying salmonella, and a preparation and an application thereof, which are mainly used for solving the technical difficult problems of high omission ratio, complicated identification steps, higher cost and non-intuitive screening result of an existing salmonella separation method. The kit disclosed by the invention comprises general-purpose type non-selective enrichment broth, general-purpose type non-selective enrichment broth which is concentrated by 10 times, selenite sulfonamide green-sulfanilamide enrichment broth, a xylose lysine deoxycholate agar plate, a salmonella enzyme chromogenic agar plate, a salmonella biochemical identification tube, a hydrogen sulfide filter paper strip and an indole filter paper strip, and is characterized by further comprising improved Robert enrichment broth which comprises the following components: 13.58g of magnesium chloride, 7.2g of sodium chloride, 4.5g of papain digestion soybean peptone, 1.26g of KH2PO4, 0.18g of K2HPO4, 36mg of malachite green and 1000ml of distilled water. The kit disclosed by the invention is mainly designed for synchronous separation and identification of salmonella (including typhoid, paratyphoid and non-typhoid and paratyphoid salmonella) in enteric pathogenic bacteria.

The invention relates to an application of a BAX PCR method in detection of salmonella in foods. The method comprises the following steps: step 1, putting a sample into bacterium enrichment broth to prepare an initial suspension; step 2, adding the bacterium enrichment broth extracted in the step 1 into the brain heart infusion broth, and carrying out secondary bacterium enrichment and re-culture;step 3, adding bacterium enrichment broth subjected to the secondary bacterium enrichment and the re-culture in the step 2 into a lysis tube, adding a protease-containing lysis solution into the lysis tube, heating the lysis tube in a heating tube, cooling the lysis tube in a cooling block, and taking and putting a lysis product into a PCR tube; and step 4, carrying out computer detection on thePCR tube in the step 3. The BAX PCR method provided by the invention is applied to detection of salmonella in foods, and rapidly carries out salmonella detection with a large batch of samples.

The invention discloses a method for monitoring salmonella. The method for monitoring salmonella comprises the following steps that an appropriate amount of a sample to be detected is mixed with a BWPsolution, the mixture is ground/stirred to obtain a suspension/mixed solution, insoluble matter is removed by centrifugation, then the bacteria is washed with normal saline, lytic cells are boiled, asupernatant is taken through centrifugation, the supernatant is placed in a sterile test tube, enrichment culturing is performed in an incubator for 8-10 hours at 36 DEG C, a BPW liquid culture is inoculated into selective enrichment broth SC of the salmonella, an enrichment culturing solution is inoculated on a SS medium in a one-ring mode, and medium-sized, circular colonies with black centerson the SS medium are selected for pure culture, and the purely cultured colonies are simultaneously subjected to dual PCR and biochemical identification. The detection time of the salmonella is shortened, the detection process is accelerated, the operation complexity is reduced, the labor intensity is reduced, the detection cost is reduced, and fast, simple, and sensitive detection of a target object is achieved, the dual PCR and biochemical identification are simultaneously carried out, so that the accuracy is improved.

The invention discloses a rapid micro-droplet type digital PCR detection method of Escherichia hermannii. The method includes the steps of extracting the genomic DNA of a to-be-detected enrichment broth sample and performing dilution for later use; designing a specific primer and probe by utilizing the conserved DNA sequence of the genome of Escherichia hermannii; and using the primer and probe toperform micro-droplet type digital PCR amplification and detection on the genomic DNA of the to-be-detected enrichment broth sample. After the PCR amplification, each micro-droplet is detected one byone by adopting a micro-droplet analyzer, the micro-droplets having fluorescence signals are interpreted as 1, and the micro-droplets which have no fluorescence signals are interpreted as 0, and ultimately, the concentrations or copy numbers of to-be-detected target molecules can be calculated according to a Poisson distribution principle and the proportion of positive micro-droplets. Micro-droplet type digital PCR can be used to directly obtain the number of DNA molecules, so that absolute quantification can be performed on the Escherichia hermannii in the sample. The method has the advantages of high sensitivity, precise quantification, wide linear range of detection, good specificity and moderate costs.

In a first aspect, the invention discloses a Staphylococcus aureus separation method for eliminating the interference of food background miscellaneous bacteria. The separation method comprises an enrichment method and a purification method, the enrichment method uses an improved enrichment broth, and the purification method uses an improved trypsin soybean agar medium; in a second aspect, the invention further discloses an application of the Staphylococcus aureus enrichment method and purification method in the separation of Staphylococcus aureus; and in a third aspect, the invention also discloses a Staphylococcus aureus separation kit prepared by the application of the separation of Staphylococcus aureus. The Staphylococcus aureus separation method for eliminating the interference of food background miscellaneous bacteria provided by the invention effectively eliminates the interference of the miscellaneous bacteria, improves a separation rate of Staphylococcus aureus in food, has the advantages such as low detection method cost, simple operation, and high accuracy, and has broad application prospects and economic benefits.