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Methods of separating, characterising and (or) identifying microorganisms using mass spectrometry Invention relates to microbiology and specifically to methods (versions) of identifying microorganisms using mass spectrometry. The method is based on identification of a microorganism in a test sample, including from a hemoculture. The method comprises the following steps: obtaining a test sample known to contain or which may contain microorganisms; selectively lysing and solubilising non-microorganism cells in the test sample with a lysing solution having pH from about 8 to about 13 to produce a lysed sample; stratifying said lysed sample on a homogeneous density buffer in a container and centrifuging the container to separate said microorganism from other components of said lysed sample, wherein said microorganism passes through said density buffer to form a microorganism residue at the bottom of said container; analysing said microorganism residue using mass spectrometry to record the mass spectrum of said microorganism; identifying said microorganism in said residue by comparing the measured mass spectrum with reference mass spectra and/or with known or predicted masses of cellular components of known microorganisms. |
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Optical indicator for detecting biological pathogens Group of inventions refers to medicine and may be used for detecting biological pathogens in a sample. A clinical indicator unit for detecting biological pathogens comprises a carrier foldable to form two opposite sheets. The inner side of the first sheet of the carrier comprises a contact surface of the sample and an absorbent pad, while on the inner side of the second sheet, there is an indicator surface opposite the contact surface; the indicator surface contains a solvatochromic stain discolouring if a bacterial pathogen is found in the test sample. The unit also comprises a results windows on the outer surface of the second sheet with the absorbent pad engaging a stained reaction zone if the first sheet is folded with the second one. The group of inventions also refers to a version of the above unit wherein the first and opposite second carriers are the two sheets or parts of the same first carrier folded to face each other, and to a method for using the above unit for bacterial infection analysis. |
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Method of determining sensitivity of microorganisms of salmonella genus to antibacterial agents Test kind of microorganisms of the genus Salmonella is incubated, which is taken at a final concentration of approximately 1000 cells/ml, with the test antibacterial agent in the known concentration previously specified for each antibacterial agent, which is optimal for exposure to the specified kind of microorganisms. Incubation is carried out at a temperature of 37°C for 24 hours. The samples are added to the aptamers specific to test species of living microorganisms, fluorescently labelled, at a final concentration of 100 nM. It is incubated for the second time for 20 minutes. The sample is examined using flow cytometry. The level of fluorescence of aptamers is determined by graphs made with a flow cytometer using a program which enables to display a percentage value. The part of bound and un-bound bacteria with the aptamers fluorescently labelled is determined. The amount of the living or non-living organisms in the sample is determined. |
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Test-system for differentiating species and biotypes of bacteria of genus yersinia Invention relates to field of microbiology and deals with test-system for differentiation of species and biotypes of bacteria of genus Yersinia. Claimed invention consists of a set of nutritional media, which contain substrates and reagents for determination of presence in microorganism of lysine decarboxylase, ornithine decarboxylase, tryptophan deaminase, urease, acetoin production, indole production, meliobiose fermentation, rhamnose fermentation, saccharose fermentation, sorbite fermentation, lipase, xylose fermentation, maltose fermentation, α-methyl-D-glucopyranoside fermentation, salicin fermentation, sorbose fermentation and raffinose fermentation. |
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Nutrient medium for extraction of bacteria of shigella kind from water objects Nutrient medium is proposed for extraction of Shigella kind bacteria from water objects. The nutrient medium contains the following components: 0.1% alcohol solution of bromcresol green - 9.0 ml; 0.2% alcohol solution of methyl red - 3.0 ml; fermentative peptone - 0.5 g; extract of fodder yeast - 4.5 g; potassium dihydrophosphate - 8.7 g; caustic soda - 1.4 g; sodium chloride - 5.0 g; distilled water - up to 1000 ml; medium pH is 6.70-6.85. |
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Method includes sample preparation, DNA isolation, PCR statement. At that in carrying out PCR, the oligonucleotide primers are used, which are complementary to sequences of the chromosomal genes fliC and hom2, having the following sequences: fliC-F: 5'-TGGAGCAGTAACAATTGG-3', fliC-R: 5'-GCACCACTGATAGAAATGTTAG-3', hom2-F: 5'-GACGTGTTAAAAGAAGCCCA-3', hom2-R: 5'-CACCAATTTCGTCTTTTACA-3', followed by electrophoretic analysis of the amplification products, when the formation of the amplification product is 153 bps in size it is indicative of belonging of the strain under study to the species B.anthracis, formation of the amplification product with size of 550 bps is indicative of belonging of the strain under study to the other species of the genus Bacillus. |
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Differential diagnostic nutrient medium for identification of yersinia bacterium Invention refers to biotechnology, microbiology, and concerns the recovery and identification of pseudotuberculosis and intestinal yersiniosis agents (Y. Pseudotuberculosis and Y. Enterocolytica). A nutrient medium contains microbiological agar (dry), lactose, glucose, urea, calcium chloride, 1% alcoholic phenol red, 1% alcoholic methylene blue and distilled water in specific proportions. |
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Method for preparing microgravimetric immunosensor Invention refers to biotechnology and microbiology. What is presented is a method for preparing a microgravimetric immunosensor. A surface of a quartz crystal resonator is activated first by plasma spray coating with polyethylene imine of molecular weight less than 10000 Da for 10 s in a vacuum unit at UHF field power 30 Wt. Thereafter, immunoglobulins from a solution with the protein concentration of 0.125 mg/ml are immobilised on the activated surface of the quartz crystal resonator. That is followed by washing in distilled water and drying in an air flow. |
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Method for quantitative assessment of bactericidal activity of disinfectants Invention refers to microbiology and biotechnology. Analysed bacterial strains are inoculated on a dense nutrient medium. Paper disks impregnated with a disinfectant are applied. They are incubated in a temperature chamber until the bacteria start growing. The bacterial growth inhibition areas are measured. A quantity of the grown colonies is counted to construct a dependence diagram of the bacterial growth inhibition area and the quantity of the grown colonies after the disinfectant reaction. The diagram and Shughart inspection sheet are used to assess the disinfectant activity on specific types of the microorganisms. The disinfectants with mean measurements of the bacterial growth inhibition area are above an upper control limit of the Shughart inspection sheet are considered to be high bactericidal activity agents. The disinfectants with mean measurements of the bacterial growth inhibition area are below a lower control limit of the Shughart inspection sheet are considered to be low bactericidal activity agents. The disinfectants with mean measurements of the bacterial growth inhibition area between the control limits of the Shughart inspection sheet are considered to be mean bactericidal activity agents in relation to all analysed agents. |
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Method of species identification of l.casei/paracasei, l.fermentum, l.plantarum and l.rhamnosus Invention refers to a method of species identification of L.casei/paracasei, L.fermentum, L.plantarum, L.rhamnosus lactobacilli. The proposed method involves performance of a PNR reaction with species-specific primers; besides, primers are built, which are specific to the first gene of operon FIFO ATP of synthase (a gene of subunit a) and a gene of uracylphosphoribosyltransferase, which precedes it, for L.casei/paracasei and L.rhamnosus and a gene of uracyltransport protein for L.plantarum and L. fermentum. |
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Dry chromogenic feed medium for detection of coliform bacteria and e.coli (versions) Invention can be used for detection of coliform bacteria and E.coli in specimens of food products and water at performance of bacteriological tests. Feed medium includes a nitrogen source represented by meat peptone or pancreatic hydrolysate of fish flour, sodium chloride, dibasic sodium phosphate, potassium monophosphate, sodium pyruvate, L-tryptophane, sodium dodecyl sulphate, 6-chloro-3-indolyl-β-D-galactopyranoside (Salmon - GAL), 5-bromine-4-chloro-3-indolyl-β-D-glucoronide-(X-GLUC), isopropyl- β-D1-tiogalactopyranoside (IPTG) and microbiological agar in the specified ratio. |
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Invention can be used for detection and considering of E.coli and coliform bacteria in water, food products, clinic material, etc. Feed medium contains pancreatic hydrolysate of fish flour dried with Tergitol 7 on the bases of 0.1 g of Tergitol 7 per 6 g of dry pancreatic hydrolysate of fish flour, yeast extract, 1-water D (+) lactose, bromthymol blue, sodium dodecyl sulphate, 2,3,5-triphenyltetrazolium chloride, sodium carbonate and microbiological agar in the specified ratio. |
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Method of identifying vibrio bacteria Invention relates to microbiology and biotechnology. Material to be investigated - pure culture of rod-like, gram-negative, glucose-fermenting, oxidase-positive or oxidase-negaive bacteria - is collected first. The investigated daily bacterial culture is seeded on the surface of nonselective nutrient agar (GRM-agar) with 1% sodium chloride. A paper disc is then placed seeded surface, said disc containing vibriostatic substance niclosamide (2,5-dichloro-4-nitrosalicylanilide) in amount of 10 mcg or 16 mcg per disc. The seeded material is incubated in aerobic conditions at 35°C for 24 hours. Vibrio bacteria are indicated a zone of inhibited bacterial growth around the disc. |
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Invention pertains to the method for quick growth, detection and identification or counting of microcolonies of microorganisms at early stage. The described method includes the following stages: obtaining the container with medium with porous element located above or under the top surface of the medium, note that the medium has nutritious substances for quick growth of microcolonies of microorganisms and porous element has pores from 1000 to 107 Da; pouring the liquid sample without serial dilution into container to the area above porous element; capturing the microorganisms in porous element or at the medium above porous element; incubation of container for the time sufficient for quick growth of microcolony at an early stage; transfer of porous element and any medium above it from container to the secondary medium for evaluation, detection and identification of microorganisms; and microcolonies research relatively the growth, detection, identification or counting of microorganisms. The said method for growth, detection and identification or counting of microorganisms takes not more than approximately six hours. |
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Method for increasing biocidal and therapeutic action of suspension-cream with linco-spectin Invention represents a method for increasing biocidal and therapeutic action of a suspension-cream with linco-spectin consisting in detoxification and polymerisation of linco-spectin 100 g in water 300 ml by 0.15±0.05% glutaric aldehyde 0.15±0.05% alkyldimethyl benzylammonium chloride at 38-40°C for 2-3 days. |
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Invention refers to medicine, namely gynaecology and may be used for individual selection of the preparations containing the probiotic lactic bacterial strains for effective intravaginal therapy. For this purpose, vaginal epithelial cells are recovered from the patient, released from the accompanying microflora. That is followed by preparing an epithelial cell suspension in a culture medium, and mixed with a suspension of thermally-activated probiotic lactic bacterial strains. Then, the suspension is incubated; the epithelial cell culture fluid filtrate is prepared and added to the suspension of the tested probiotic lactic bacterial strains in ratio 1:7. Concurrently, a reference of the mixture of the epithelial cell culture medium and the suspension of the tested probiotic lactic bacterial strains in ratio 1:7 is prepared. The test and reference samples are incubated, measured for optical density; and a degree of biomass increase in the test sample is related to that in the reference. The preparation containing the probiotic lactic bacterial strains the biomass increase of which under the influence of patient's vaginal epithelial cells is stimulated most is selected form the effective intravaginal therapy. |
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Method of determining genus of bacteremia agents Method comprises incubation of the blood sample in the nutritional medium, followed by detection of microbial growth in the primary hemoculture. Sample preparation of the sample under study is carried out by centrifugation. The quantitative chromatography-mass-spectrometer analysis of the sample under study is carried out with the definition of marker molecules, which are used as molecules of free and substituted higher fatty acids of cellular lipids of microorganisms. The free and substituted higher fatty acids are identified by comparing the data obtained with the database NIST of chromatography-mass-spectrometer. The genus of bacteremia agents is determined using the Table 1. |
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Method of determining minimal inhibitory concentration of antimicrobial preparation Microorganism-causative agent is isolated from the biological material of the patient. The standard suspension of the isolated microorganism-causative and two-fold dilutions of antibacterial preparations with known activity is prepared. Inoculation is carried out. The standard suspension of the isolated causative agent with the volume of 0.1 ml is mixed with 5.0 ml of liquid nutrient medium specific for this pathogen. It is incubated for 24 hours at +37°C. The test 1:100 and control 1:200 dilutions are prepared in the same nutrient medium. Each of the two-fold dilutions of antibacterial preparations in amount of 0.075 ml with an equal amount of the test dilutions is applied in the test cavities of the plate. In the control cavity 0.150 ml of control dilution is applied and incubated for 48 hours at + 37°C. The liquid fraction of the suspension is removed from the cavities. For staining 0.2 ml of 1% solution of crystal violet is added to each cavity. It is exposed at room temperature for 30 min. The contents of the cavities are washed three times with distilled water and 0.2 ml of 99% solution of dimexidum is added to each cavity. It is exposed at room temperature for 15 minutes. The optical density of the medium in the control (ODC) and the test cavities is measured in the optical units (pu) in a microplate reader at the wave-length of 540 nm. When fulfilment of the condition ODC> 0.2pu, the greatest two-fold dilution of antibacterial preparation in the test cavity with an optical density less than 0.2 pu is defined as the minimum inhibitory concentration (MIC). |
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Method of detecting mycobacterium tuberculosis in air environment Invention relates to microbiology and is meant for detecting mycobacterium tuberculosis in the air environment of prevention and treatment facilities. The method involves collecting air samples through directed impact action of a jet on a foam plate in a Petri dish, said plate being soaked in 4-6 ml of 5% trisubstituted sodium phosphate solution followed by incubation of the Petri dish wit the foam plate for 18-24 hours; centrifuging the analysed material, followed by separation of supernatant fluid and neutralising the precipitate with 1% citric acid solution in ratio of 1:1, followed by inoculating the neutralised precipitate on a Novaya culture medium and culturing until growth. The diameter of the foam plate is equal to the diameter of the bottom of the Petri dish and its thickness is 4-6 mm. |
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Method of detecting and counting viable legionella pneumophila microorganisms Present invention relates to microbiology and a method of detecting and counting viable Legionella pneumophila microorganisms in a sample. The described method involves: (1) contacting said microorganisms of said sample with at least one reducing compound which contains glutamate and pyruvate, and a culture medium which is a buffered charcoal yeast (BCYE) or GVPC agar culture medium, (2) incubating the product of step (1), and (3) detecting and determining the number of viable microorganisms. The reducing compound used directly or indirectly affects metabolism, reducing oxidative stress of the microorganism by reducing formation and/or breaking down reactive forms of oxygen. |
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Invention may be used to assess biological activity of lactobacilli and bifidus bacteria relative to Vibrio cholerae with the purpose to establish possibility to use probiotics for prevention and treatment of cholera. The method provides for testing of antagonistic and acid-forming activity of lactobacilli and bifidus bacteria strains. Antagonistic activity relative to choleraic vibrios is detected by the method of holes in dense nutrient media - CDM - agar and alkaline agar, and activity is accounted on the basis of quantitative data, namely: zone of growth inhibition (in mm) of a test strain V. cholerae from 6.0 to 15.0 - low antagonistic activity, from 15.1 to 29.0 - medium, ≥29.1 - high. Acid-forming activity of strains relative to choleraic vibrios expressed in Turner degrees (°T) is defined on the basis of quantitative data. So if acid formation ≤is 99.9 (°T), then activity of acid formation is assessed as low, if the values of this index are within (100-149.9) (°T) - as medium, and if the value is ≥150.0 (°T) - as high. At the same time the result of assessment of lactobacilli and bifidus bacteria biological activity relative to V. cholerae eltor, classica, O139, non O1/ non O139 is considered positive with 4 versions of combination of testing results according to the specified indices, namely: high antagonistic activity, high acid-forming activity, accordingly, high - medium, medium - high, medium - medium. |
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Method of diagnostics of candidiasis of upper respiratory tract in workers of agroindustrial complex Method provides sampling of the material under study. Inoculation of the material under study on nutrient medium sabouraud followed by incubation on this medium at 37°C for 24 hours. A quantitative assessment of growth of yeast-like fungi of the genus Candida, and with a value exceeding 10*1 CFU/swab the candidiasis of upper respiratory tract is diagnosed. The obtained colonies are examined under the microscope and the isolated colonies of yeast-like fungi of the genus Candida are separated from these colonies. The isolated colonies of yeast-like fungi of the genus Candida are suspended in five test tubes containing liquid medium sabouraud with the phenol red indicator, where the discs with carbohydrates are added, and in the first test tube - a disc containing maltose, in the second - sucrose, the third - lactose, the fourth-galactose, the fifth - trehalose, and are incubated at 37°C for 24 hours, followed by assessment of colour change of the indicator. The colour change of the indicator to yellow is taken as one, lack of colour change is taken as zero, the sum of the values obtained in assessment of the indicator colour in five test tubes is calculated. If the value of the sum is 0 the candidiasis of upper respiratory tract caused by Candida kruzei is diagnosed, if it is 1 - the candidiasis of the upper respiratory tract caused by Candida glabrata is diagnosed, if it is 3 - the candidiasis of the upper respiratory tract caused by Candida albicans is diagnosed, if it is 4 - the candidiasis of the upper respiratory tract caused by Candida tropicalis is diagnosed, if it is 5 - the rest. |
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Elective-differential nutrient medium for extraction of choleraic vibrios Invention is an elective nutrient medium, which may be used in laboratory practice for extraction of a cholera agent. The nutrient medium contains a nutrient base, microbiological agar, saccharose, sodium chloride, sodium carbonate, bismuth nitrate pentahydrate, sodium citrate, cattle bile, EDTA iron (III)-sodium salt, bromthymol blue, mesoinositol, activated charcoal, sodium sulfite and L-cysteine, potassium tellurite and distilled water. The nutrient base in the nutrient medium is a dry nutrient broth from a pancreatic overcook of Caspian sprat or a dry nutrient broth from hydrolysate of fish and bone flour or a complex of yeast extract and peptone at the specified ratio of components. |
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To assess effect of exogenous factors at intestinal microflora in case of its dysbiotic disturbances in an examined individual they determine the quantity of microorganisms of endogenous flora per 1 g of faeces before start and upon completion of exogenous factor effect. The parameter of endogenous flora microorganism quantity variation speed is calculated in the form of their content per 1 g of faeces per day by calculation of the difference of values of endogenous flora microorganisms quantity per 1 g of faeces between start and end of examination and subsequent division of this difference into amount of days that elapsed between measurements. The produced parameter is compared with the average statistic parameter of endogenous flora microorganism quantity variation speed per 1 g of faeces, calculated in a advance on a large group of patients with a similar pathology of gastrointestinal tract. On the basis of comparison results they conclude on efficiency or no effect of exogenous factors in case of dysbiotic disturbances of intestine. |
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Method according to the invention discloses the possibility to detect β-lactamase-producing oxidase-positive gram-negative diplococci, suspicious for appurtenance to Moraxella (Branchamella) catarrhalis and provides for inoculation of a clinical material on blood-serum agar in sectors with simultaneous application of discs with penicillin 10 AU. The inoculation is incubated under regular atmospheric conditions for 18-24 hours, and in 36-48 hours from the moment of inoculation they assess availability or absence of a zone of growth delay for those microorganisms that are present in the clinical material. If there is no zone of microorganisms growth suppression around discs, they detect availability in the clinical material β-lactamase-producing oxidase-positive gram-negative diplococci. Appurtenance of detected diplococci to Moraxella (Branchamella) catarrhalis is identified by biochemical criteria. |
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Nutrient medium contains HMF - agar, erythrocytic mass from donor's human blood, serum of cattle and yeast extract at the specified ratio. |
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Method to monitor microbiological activity in technological flows Device comprises a through cell equipped with holes, where at least one hole represents an inlet hole for intake of fluid medium from the specified technological flow, and at least one hole is an outlet hole for discharge of fluid medium from the specified through cell. To one of specified holes an RK probe is attached, possibly, an OVP probe, a cleaning accessory. The first pipeline is connected to the inlet hole. Possibly, the second pipeline is connected to the outlet hole. A valve is connected to the specified through cell. With the help of the specified devices and methods they measure volume microbiological activity and surface microbiological activity in a process flow of water by means of measurement of dissolved oxygen concentration. |
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Method includes the following stages: contact of a sample with a source of nutrition for cells, containing antioxidant, representing pyroracemic acid or its salt, and an inhibitor of cell proliferation, which is selected from ciprofloxacin and cefalexin; contact of the specified sample with fluorescent-marked oligonucleotide probes, capable of specific hybridisation at least with one section of ribosomal nucleic acids, which belong to microorganisms of Legionella pneumophila kind and type; and detection and quantitative determination of a fluorescent signal. |
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Method under the invention provides detoxification and polymerisation of antibiotics in 0.15±0.05% glutaric aldehyde at 38-40°C for 3-5 days, and then in 0.1% alkyl dimethyl benzyl ammonium chloride at 38-40°C for 3-5 days. |
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Method for enhancement of efficacy of e.coli resistant antibiotics Method under the invention involves detoxification and polymerisation of the tested antibiotics 150-200 mg/ml at first in 0.15±0.05% glutaric aldehyde at 38-40°C for 2-3 days, then in 0.1% aethonium or 0.1% alkyl dimethyl benzyl ammonium or 0.1% Biopague D at 38-40°C for 2-3 days. |
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Thermostatting of a biocatalyst is carried out on the basis of immobilised microbial cells and a non-innoculated carrier within the biocatalyst, as well as infrared scanning of the biocatalyst surface and the carrier with the help of a highly sensitive infrared chamber, and production of thermal characteristics of the biocatalyst, such as distribution of temperatures on its surface and difference of temperatures between the surface of the biocatalyst and the non-innoculated carrier. Distribution of temperatures makes it possible to control homogeneity of activity distribution on the biocatalyst surface. The difference of temperatures is used to determine intensity of adsorption and metabolic activity of fixed bacterial cells. A plant for detection of efficiency of adsorption immobilisation of microorganisms and monitoring of functional condition of biocatalysts includes an infrared chamber fixed on a tripod, and connected with a computer, and a heat-insulating box with a hole on top, closed with a cover, which makes it possible to minimise oscillations of ambient temperature down to ±1°C/hr and reduces impact of the infrared chamber at analysis results. |
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Method for differentiation of strains of vibrio cholerae serogroup 0139 by alkyl sulphate activity Invention refers to medical microbiology and concerns differentiation of toxigenic and non-toxigenic strains of cholera vibrios serogroup 0139. The described method involves preparing a synthetic medium 100 ml by using weights of: sodium chloride 0.5 g, agar 2 g, bromthymol blue 0.002 g; then the weight ingredients are dissolved in 0.01 M tris HCl buffer 100 ml, pH 7.8 and boiled for 30 minutes; the prepared medium is added with a substrate in the form of 1% aqueous sodium dodecyl sulphate (SDS) to the final concentration of 0.1% in the medium; it is followed by loop inoculation of the prepared medium on the analysed culture and incubation for 24-48 hours; the results are considered by the presence of milky-white areas 2-10 mm surrounding the inoculations on the agar sectors; the presence of those makes the strain to be referred to as toxicogenic (ctx+ tcp-), while the absence of those shows that the strain is non-toxicogenic (ctx+ tcp-). |
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Method of isolating uncultivable staphylococcus bacteria Invention relates to field of medicine, namely to microbiology and is intended for increasing efficiency of microbiological diagnostics of staphylococcal infections. Method of isolating uncultivable staphylococcus bacteria includes cultivation of studied material on beef-extract agar for 24 hours at temperature 37°C. After that, grown bacterial culture is kept at temperature +4°C for 48 hours. |
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Culture medium for determining drug susceptibility of m tuberculosis to main anti-tuberculosis drugs Culture medium for determining drug susceptibility of Mycobacterium tuberculosis to four main drugs - streptomycin, isoniazid, rifampin and ethambutol contains: 4.7 g dry Middlebrook 7H9 broth, 1.25 g microbioligical agar-agar, 1.25 g pancreatic digest of casein, 900 ml distilled water and 100 ml growth additive OADC. The additive contains, g/l: sodium chloride 8.5, bovine albumin (fraction 5) 50.0, glucose 20.0, catalase 0.03 and oleic acid 0.6 ml/l. |
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Method of detecting antibodies to mycobacterium leprae on solid carrier Claimed is method of detecting antibodies to Mycobacterium leprae (M.leprae) on solid carrier. Places of application of components of immunologic reaction in ELISA on solid carrier (fluoroplastic tape) are sensitised with antigen from ultrasound disintegrate (sonicate) of M.leprae in dose 5 mcg/ml in volume 20 mcl for each sample of patients' blood serum. After that immunoperoxidase conjugate against human IgG is applied in the same volume on the places of previous location of reaction components. Unbound antigen from sensitised tape is removed with buffer solution (BPST) after each stage of reaction. Reaction results are estimated visually by difference in substrate mixture colouring. |
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In in vitro conditions, imitating digestion process in humans quantities of live microorganisms at the beginning and at the end of experiment are determined and their numeric values are compared. Conclusion is made on the basis of the results of comparison after incubation of probiotic microorganisms during 4 h in acidic model medium with acidin-pepsin by inoculation of microorganism suspension on dense nutritional medium. Grown colonies are counted and number of viable microorganisms is determined. Remaining suspension is separated from incubation medium, alkaline model medium with pansinorm forte 20000 is added to sediment in volume, analogous to volume of acidic model medium. Sediment is resuspended and suspension is incubatred during 12 hours, remaining number of viable microorganisms is determined by inoculation on a dense nutritional medium and counting grown colonies. |
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Diagnostic technique for microsporia Diagnostic technique for microsporia in implemented by featuring clinical manifestations of the process, studying macro-and microphology of the culture. The presence of 1-2 centres of size 0.5-1.5 cm primarily localised on the scalp and the presence of two individual cell macroconidia in the form of a horseshoe with a thick pitted wall enable diagnosing microsporia caused Microsporum cams var.distortum. By using the complex of anamnestic, clinical, microscopic and morpho-biological signs, the invention allows establishing a precise etiological diagnosis of a microsporia agent - M canis var. distortum that is important in the process of the antimycotic therapy. |
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Method for recovering pure staphylococcus culture Method provides grinding a pathological biomaterial to be homogenated to prepare a suspensions. The prepared suspension is added with 3-5% citric acid at the basis of its content in the suspension. It is kept for 20-30 minutes at 10-20°C and added with 3-4% succinic acid and kept for 20-30 minutes at 10-20°C that is followed by neutrilising the suspension with 5-10% ammonium and reducing to pH 7.5-7.6. The neutrilised suspention is settled for 20-30 minutes; a supernatant is rinsed, and the precipitation is inoculated with a nutrient medium containing in 1 l of distilled water citric acid 8.0 g, ammonium citrate 2.0 g, succinic acid 3.0 g, asparagine or glycine 2.0 g, di-basic potassium phosphate 5.0 g, magnesium sulphate 0.5 g, zinc sulphate 0.3 g, di-basic sodium phosphate 3.0 g, sodium chloride 5.0-6.0 g, ferrous sulphate 0.1 g and glycerin 40-50 ml at pH 7.5-7.6. To produce the solid agar medium, the fluid medium diluted in distilled water 1:1 is added with agar 2.5 g per the fluid medium 100 ml, and sodium chloride is reduced to 5-6%. |
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Technique consists in total determination of the content of ammonium, amines and easily volatile carboxylic acids in an equilibrium gas phase over patient's cervical mucus with using an 'electronic nose' detector. The electronic nose has a package of 8 piezoelectric sensors and connected to a computer for data processing. Each sensor has a basic vibration frequency 10-15 MHz. The electrodes are coated with films of acetone solutions of bromocresol blue, methyl red, triton X-100, toluene solutions of polyethylene glycol 2000, polyethylene glycol adipinate, dicyclohexane-18-crown-6 and bee wax, and a chloroform suspension of multilayer carbon nanotubes. Total weight of each coating after solvent removal is 4-10 mcg. The technique is enabled by applying the equilibrium gas phase 1 cm over patient's cervical mucus in the cell of the device. The package of the piezoelectric sensors generate signals to be registered in the form of chronoperiodograms, the sorption parameters of ammonium, amines and easily volatile acids. The parameters are used to form a special data array and processed. The result is described as 'healthy' or 'sick' with specifying the disease: clamidiosis, candidiasis, gardnerellosis, trichomoniasis, ureaplasmosis, mycoplasmosis or their combination. The presence of pathogenic microorganisms in the cervical mucus sample is conditionally acceptable as +1, while the absence is coded as (-1) (reference values). |
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According to the first version, vaginal epitheliocytes are produced, separated from accompanying microflora, incubated in the Hanks' medium with added cefuroxime and lisocyme; a supernatant is separated. A microbial suspension is prepared from the daily culture of the analysed bacteria and mixed with the epithelial supernatant. It is combined with preparing a control suspension of the analysed bacteria and an epitheliocyte cultivation medium, incubated; the biological properties are measured in the bacteria of the test and control samples. A degree of the varying biological properties in the test samples is evaluated by the relation to the control sample, and the degree enables stating stimulation, indifferent or supression action of epitheliocyte on the biological bacterial properties. According to the second version - the vaginal epitheliocytes are produced, inactivated by formalin, washed out to prepare an epytheliocyte suspension in physiologic saline. A microbial suspension is prepared from the daily culture of the analysed bacteria and mixed with the inactivated epithelial suspension. It is combined with preparing a control sample from the analysed bacterial suspension and physiologic saline; the test and control samples are incubated; the biological properties are measured in the bacteria of the test and control samples; a degree of the varying biological properties in the test samples is evaluated by the relation to the control sample, and the degree enables stating stimulation, indifferent or suppression action of epitheliocyte on the biological bacterial properties. |
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Clinical effectiveness is estimated on an simulation model and provides creating a model of bacterial biofilm grown on biolumenescent bacteria Vibrio fischeri. The biofilm is exposed to antibiotics and ultrasound to estimate an antimicrobial effect. The bacterial viability variation may be controlled by varying luminous intensity of the biolumenescent bacteria with using devices, e.g. lumen meters. The antimicrobial effect is estimated by a degree of luminous intensity inhibition as compared to the reference. |
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Method provides DNA recovery of an analysed plague agent strain that is followed by conducting a PCR with the use of oligonucleotide primers for streptomycin, chloramphenicol and kanamycin resistance genes - strA, strB, cat and npt (nucleotide sequences of the primers are presented in the description). The antibiotic-resistance genes are detected by the presence of the amplified fragments of the strA, strB, cat and npt genes having the sizes of 387, 705, 377 and 361 base pairs respectively. |
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Method for recovering pure culture of tuberculosis mycobacteria Pieces of the affected organs (a biomaterial) are grinded to prepare a suspension. The prepared suspension is filled with a solution containing 4-6% of citric acid and 3% of hydrogen peroxide in the ratio 1:1 to the biomaterial suspension and kept for 30-60 minutes at room temperature to be thereafter processed in a solution containing 4-5% of succinic acid and 3% of hydrogen peroxide in the ratio to the suspension 1:1. It is kept for 30-60 minutes and then neutralised with 5-10% ammonia to pH 7.0. It is followed by decantation of the supernatant, and the Lowenstein-Jensen agar medium is inoculated with the precipitation. |
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Method for selection of mycotoxin decomposer microorganisms for prevention of mycotoxicosis For the purpose of selection of mycotoxin decomposer microorganisms for prevention of mycotoxicosis, the examined culture is inoculated on a selective nutrient lactobacagar medium containing mixed mycotoxins and resazurin, and a mycotoxin-free test medium, and cultivation. After the cultivation is completed, microorganisms are examined for their ability to metabolise toxic metabolites by decolouration of intensive red colour to colourless and to detect the mycotoxin decomposer microorganisms by the diametre of a decolouration area, maximally close to the reference. |
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Method of evaluating bacterial load of urine, prostatic secretion, and ejaculate Bacterial load of substratum load is evaluated by sampling a material, taking its culture on selective nutrient mediums, recovering and identifying cultures. Urine, prostatic secretion, ejaculate cultures in sequential dilutions 10-1 to 10-10 are taken on Blaurock medium, Schadler broth, Schadler agar, blood agar (BA) on the basis of Muller-Hinton medium, bile-esculin agar for bacteroids - for cultivation of non-clostridial anaerobic bacteria. It is also combined with taking on Endo medium, chromogenic selective agar for enterococci, chromogenic selective agar for candida, chromogenic selective agar for Klebsiellas, chromogenic selective agar for staphylococci, BA on the basis of Muller-Hinton medium - for cultivation of optionally anaerobic bacteria. The optionally anaerobic and non-clostridial bacteria are detected with evaluating its concentration from minimal dilution level 10-1. |
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Method of evaluating bacterial load of urine, prostatic secretion, ejaculate Method is enabled by taking urine, prostatic secretion, ejaculate cultures on solid mediums, in addition taking substrata cultures on Blaurock semi-solid medium and Schadler broth in sequential dilutions 10-1 till 10-10 to detect non-clostridial anaerobic bacteria. |
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Method for differentiating francisella tularensis subsp mediasiatica bacteria Invention relates to medical microbiology and a method of differentiating Francisella tularensis subsp. mediasiatica bacteria. The method involves testing the analysed culture using an indicator and a substrate in form of discs with nitrocefin. The discs are first washed with distilled water and then placed on the analysed strains. Further, incubation is carried out at 37°C for 5-10 minutes. Results are determined from the colour of the discs. Francisella tularensis strains of the Central Asian subspecies do not change the colour of the discs, while strains of other subspecies colour the discs red. |
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Method of detecting microorganisms in sample Invention relates to microbiology. Biological or artificial fluid medium containing the selected microorganism is centrifuged. Supernatant fluid is filtered. A series of diluted samples corresponding to increase in filtrate dilution of up to 10-15 is obtained. The samples are exposed to an electric, magnetic and/or electromagnetic excitation field. Electric signals detected by a solenoid and the digital record of said electric signal after passing through an analogue-to-digital converter are analysed. Diluted samples for which characteristic electric signals, whose amplitude is 1.5 times higher than that of the background noise signals emitted by water, are obtained are selected. Test tubes with equal volume of diluted samples are placed in protective jackets to protect the diluted solutions from external electromagnetic fields. The solution contained in test tube T1 is used as the standard solution. Test tube T2 is placed in the immediate proximity of the sample or is brought into contact with sample X which is presumed to contain the selected microorganism (e.g., E.coli). The obtained electromagnetic signals are compared. Suppression of the signal indicates presence of the microorganism in sample X. |
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Method for standartisation of control seed strain pasteurella multocida Invention concerns a method for standartisation of control seed strain of chicken cholera agents (Pasteurella multocida). The presented method involves cloning of the S- or M-form Pasteurella strain, gradual passaging by intramuscular, and then intranasal introduction in a susceptible organism of weight about 350 g; in 3 hours following after the death of every infected body, the microorganism is respectively isolated into physiological saline, and sealed by into 0.5-1.0 ml Pasteur pipettes with the Pasteurella strain isolated by the intramuscular passage being used over a period of 15-20 days as a by-product for the intranasal passage, while the Pasteurella strain isolated by the intranasal passage is used 2-3 days following the isolation procedure during the next 3 days as a reduction for producing a 10-hour (9 h at 37 and 1 h at 20°C) broth culture of encapsulated Pasteurellas. |
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Method of producing culture medium for investigating microbial contamination of air Dry enzymatic peptone, glucose, dialysate of baker's yeast and microbiological agar are mixed in given quantities. The obtained mixture is added to 1 litre of distilled water and boiled until complete dissolution of the agar. While hot, the mixture is poured into vials, sterilised in an autoclave and cooled to 47-55°C. The cooled medium is poured into Petri dishes and held until complete setting of the gel. Weighed zinc nanopowder in a physiological solution in amount of 0.005 mcg of zinc nanopowder per 0.1 ml of the physiological solution per Petri dish is deposited on the solid surface of the gel in aseptic conditions. |
Another patent 2513513.