Identification method of blue pus bacillus pseudomonas aeruginosa |
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IPC classes for russian patent Identification method of blue pus bacillus pseudomonas aeruginosa (RU 2540501):
   
 Method of analysis of removal of biochemicals with meadow grass / 2538802
For analysis of removal with meadow grass of biochemicals the yield fluctuations are accounted depending on the structure of phytocenosis in the form of a ground cover. Conducting statistical data processing of testing the samples of grass from the test plots on the meander, central and terrace near flood plains, and also on the meadows with uneven and tiling location of grass species. And prior to establishment of sample plots the terrain reconnaissance is carried out with the grass cover selected for the measurements, an outline map of location of the components of the grass cover is made, then on each component of the grass cover in the form of a plot at least one temporary test plot is established, the wet and air-dry weight of the sample is determined by weighing on the cut grass sample. At that the tests on biochemical analysis are additionally carried out on the dried samples of grass to determine the concentration of at least three chemical nutrients in the form of mobile nitrogen, potassium and phosphorus oxides, and then by summing the concentrations of these three substances the total summarised removal of substances from the aerial part of grass on all test sites is calculated.
 Method for confirmation of death caused by ventricular fibrillation accompanying myocardial infarction / 2538622
For the purpose of the confirmation of a patient's death caused by ventricular fibrillation accompanying myocardial infarction in the patients died from myocardial infarction in the course of autopsy on the basis of a macroscopic pattern, the pathological process age is determined. This enables selecting examination areas in the heart at the level of necrosis. Absolute values (AV) of the following cell populations are counted in the x600-magnified visual field with using haematoxylin and eosin: necrosis-borderline lymphocytes (Lfb) in myocardial infarction of the age of 1-2 days, necrosis neutrophilic granulocytes (NGn) and fibroblasts (Fbn) in myocardial infarction of the age of 3-5 days. The derived absolute values are compared to threshold values (TV) with the threshold values of Lfb=2, NGn=38, Fbn=1. Each visual field is assigned with a diagnostic coefficient (DC), which is equal to 1.9 at the age of myocardial infarction of 1-2 days, if the absolute values Lfb<2 or -3.9, Lfb>2; if the age of myocardial infarction is 3-5 days, the diagnostic coefficient is -2.1, if AFbn<1, or the diagnostic coefficient is 7.2, if the absolute value of Fbn>1; the diagnostic value of 1.36 is shown by the absolute value of NGn<38; the diagnostic value is -11, if the absolute value is NGn>38. The total value (ΣDC) is calculated by moving between the visual fields within the examined area, and if ΣDC>12.78, the patient's death is stated to have come from ventricular fibrillation; if ΣDC<-12.78, the death is stated to be caused otherwise.
 Device and method for microbiological analysis of biological samples / 2531047
Invention refers to laboratory diagnostics and can be used for testing biological samples for pathogenic microorganisms. The device for microbiological analysis of the body fluid samples comprises a compartment for incubating sample containers, an analyser for studying the internal atmosphere of the above containers and a sorting system for classifying the above containers in accordance with the carbon dioxide content detected by the above analyser. The sorting system classifies the containers delivered from the above incubation compartment to divide the above containers into the positive containers to be analysed for identifying microorganisms found in the respective samples, and the negative containers wherein the analysis is not required for identifying the microorganisms, and also the unspecified containers to be incubated once again.
 Method for determining clinical effectiveness in anti-inflammatory process of septic wounds under physiotherapeutic exposure / 2530630
Method involves a post-therapeutic cytological examination of smears sampled from septic wounds, a cell composition analysis, including neutrophilic granulocytes and tissue elements - fibroblasts and fibrocytes; the material sampled from the septic wound on the 3rd day from the beginning of therapy is used for the cytological examination to determine accessory signs of therapeutic pathomorphism, namely the morphology of neutrophilic granulocytes, fibroblasts and fibrocytes and the presence of oxyphilic ground substance; observing the nuclear fragmentation and cytoplasm vacuolation in the neutrophilic granulocytes, multinuclearity, nuclear lobulation, karyonuclei in nuclei, basophilia, cytoplasm grain and vacuolation in fibroblasts and fibrocytes, as well as the presence of the oxyphilic ground substance, the treatment is stated to be effective.
Diagnostic technique for renal functional reserve / 2528903
Invention refers to medicine. Substance of the early diagnostic technique for chronic renal disease consists in using a dosage range of dopamine of 1 to 3 mcg/kg of body weight and a standard water load of 200 ml. No glomerular filtration rate increase testifies to the presence of early signs of chronic renal disease.
 Method for quantitative measurement of carbon nanostructures in biological samples and body distribution thereof / 2528096
Carbon nanostructure surface is modified with (2,4,5-triiodophenyl)-methanol; the modified carbon nanostructures are analysed to measure a iodine amount; the prepared modified carbon nanostructures are administered into an experimental animal's body to take organs and tissues thereafter, to homogenise them in a 0.5-2 M NaOH solution, to sample a homogenate, to dilute with water, to expose the diluted sample to ultrasound to a temperature of 40-70°C, to determine a iodine amount in the prepared sample by mass spectrometry with inductively bound plasma and to calculate the carbon nanostructure content in the sample as shown by a difference of the iodine amount in the sample prior to administration of the modified carbon nanostructures and after administration thereof into the body and to re-calculate this iodine amount into the carbon nanostructure content in the sample with the use of the initial iodine content in the modified carbon nanostructure.
Method for prediction of clinical effectiveness of patients with non-hodgkin lymphomas with bone marrow involvement / 2526796
Invention refers to medicine, namely to oncohaematology, and can be used for prediction of the clinical effectiveness in the patients with non-Hodgkin lymphomas with bone marrow involvement. That is ensured by determining a lymphocyte volume and electric conductivity in patient's peripheral blood before the treatment and every next course of the chemotherapy with calculating a relation of the values. If observing a decrease of the relations as compared to the initial value, the positive clinical effectiveness and the absence of generalisation symptoms are predicted. The absence of the decrease or increase of the relations shows the absence or low clinical effectiveness and the presence of the symptoms of a continued tumour growth.
Method of differential morphometric diagnostics of erythrodermal form of mycosis fungoides and syndrome of skin pseudolymphoma by relative volume of epidermis and mitotic index of epidermal cells / 2526180
To perform differential morphometric diagnostics of an erythrodermal form of mycosis fungoides and the syndrome of skin pseudolymphom performed are: histological analysis of the affected skin biopsy samples by a method of light microscopy, immunodifferentiation of infiltrate cells, additional morphometric analysis of basal keratinocytes in histological preparations of the affected skin biopsy samples. In case of the similar histological picture, described in case of erythrodermal form of mycosis fungoides and the syndrome of skin pseudolymphoma such as acanthosis, hyperkeratosis, parakeratosis, oedema and vasodilatation of the derma, lymphohistiocytic infiltrates, located in the upper third of the derma, similar immunophenotype of the infiltrate cells in case of erythrodermal form of mycosis fungoides and the syndrome of skin pseudolymphoma, represented by CD2+, CD3+, CD4+, CD5+, CD43+, CD45RO+ lymphocytes, morphometric analysis of all epidermis cells is performed. After that, the relative volume of epidermis and mitotic index of the epidermal cells are calculated. In case of increase of the relative volume of the epidermis by 2.2 and more times in comparison with the normal skin and an increase of the mitotic index of the epidermal cells by 2.7 times and more in comparison with the normal skin, erythrodermal form of mycosis fungoides is diagnosed, and in case of the unchanged relative volume of the epidermis and the mitotic index of the epidermal cells in comparison with the normal skin, the syndrome of skin pseudolymphoma is diagnosed.
 Method of controlling biochemical reactions / 2525439
Invention relates to biochemistry and can be used to control biochemical reactions in vitro and in vivo. Control is carried out by exposing a magnetic nanosuspension containing a bioactive macromolecule, attached directly or through a ligand to single-domain magnetic nanoparticles, to an external low-intensity low-frequency alternating magnetic field which provides deformation and/or change in conformation of bioactive macromolecules participating in the reaction.
Diagnostic technique for arteriovenous fistula stenosis in long-term hemodialysis patients with end-stage chronic renal disease / 2522397
Arteriovenous fistula stenosis in long-term hemodialysis patients with end-stage chronic renal disease is diagnosed by four or more criteria: mean corpuscular volume (MCV) more than 93.3 ml, mean corpuscular hemoglobin (MCH) of more than 32.2 pg/l, urea concentration (Urea) more than 15.9 mmole/l, post-dialysis urea (Ur_KT/V) more than 4.2 mmole/l, mid-dialysis urea (Ur_PCR) more than 4.5 mmole/l, as well as dialysis efficacy (Kt/V) less than 1.71 units and equilibrated Kt/V (Kt/Veqv) less than 1.54 units.
 Express-diagnosis method for determining blood erythrocytes condition in fetus and newborns suffering from hemolytic disease / 2249213
Method involves examining blood erythrocytes. Blood erythrocyte suspension fluorescence intensity is measured at wavelength of 500 nm polarized along light axis and then at wavelength of 493 nm in perpendicular to the light axis for each polarization direction with following anisotropy value being calculated. Its deviation from norm proves availability of hemolytic disease.
Method for detecting endotoxicosis / 2249214
Method involves carrying out blood erythrocyte micro electrophoresis in alternating electric field with sign change frequency equal to 0.3-1 Hz. Electric field having current intensity equal to 4.5-5.0 mA is applied. Treatment is applied at 36-38°C. Oscillation amplitude being equal to 3 mm and less, endotoxicosis is detected.
Method for evaluating cardiovascular system condition in children suffering from insulin-deficient diabetes / 2250463
Method involves applying integrated diagnostics approach with blood serum examination being applied using infrared spectrometry methods. Sample absorption spectra in the area of 1200-1000 cm-1 are recorded, absorption peak heights having maximum at 1170, 1165, 1140, 1125, 110, 1070, 1025 cm-1 points are measured. Then, mean peak height C and ratio X of peak height at 1125 cm-1 point of maximum to C are calculated. X≥0.7 being the case, light myocardium injury degree is to be diagnosed. 0.7>X≥0.53 being the case, moderate severity degree case is to be diagnosed. X<0.53 indicates grave injury to take place.
Method of selecting drugs for topical treatment of gastroenteric tract diseases / 2253866
Cells of tested organs are subjected to alternating cytological electrophoresis. When cell vibration amplitude rises by at least 20%, drug is regarded to be appropriate for treatment.
Method for predicting gilbert's disease / 2253867
The present innovation deals with detecting bilirubin content both before and after curative starvation, moreover, curative starvation one should introduce antipyrine at the dosage of 1 mg/kg, in a day it is necessary to detect antipyrine content in saliva and urine and bilirubin content and at bilirubin content being 30-40 mM/l one should starve at daily intake of 400-450 kcal/d for 3 d, then one should again detect bilirubin level and at its increase by 50-100% one should again introduce antipyrine at the dosage of 1 mg/kg and according to its delayed half-life by 20-50% it is possible to diagnose Gilbert's disease.
 Method for choosing therapeutic tactics in case of chronic and acute rhinosinusitis / 2254805
The present innovation deals with treating diseases of the upper respiratory tract. One should detect disorders of mucociliary transport (MCT) due to measuring the motor parameters of ciliated epithelium cilia, at least, the rate of mucociliary transport that includes material sampling for further testing as ciliary epithelial cells and registration of cilia fluctuations in human respiratory tract due to TV microscopy performed in patient's lifetime. Material for testing should be sampled from mucosal surface of respiratory tract, and therapeutic tactics should be chosen depending upon motor velocity of cilium's tip obtained based upon mathematical modeling followed by calculations by the formula:
 Method and device for producing medical diagnostic device containing reagent / 2256167
Device has non-absorbing substrate having hydrophilic target region, which is covered with reagent by applying non-impact micro-drop printing method to produce practically uniform reagent layer. The device is in particular usable for measuring blood coagulation time. Preferential invention embodiment involves determining blood coagulation time by carrying out monitoring of light transition through the target region as the blood sample cover interacts with the reagent.
 Method for predicting functional state in bioobjects / 2258227
One should register the values of pH and Redox potential (Red) of liquid means in bio-objects simultaneously. It is necessary to plot a graph ▵pH/▵Red according to fluctuations of variation values in measured parameters at their temporal diagram to transfer it into a columnar one where the height of every column is proportional to the area of this graph between neighboring measurements, correspondingly. According to the ratio of area sums of positive columns to that of negative ones during a certain period of time ( from a minute- to a year-long ones) one should conclude upon a state in a bio-object. The value ranged 0.5-2 is considered to be a standard. The method enables to provide efficient control for a two-phase systemic process.
 Method for differential diagnostics of cerebral tumors / 2258932
One should perform crystallographic study if cerebrospinal fluid and at protein concentration in liquor samples being from 0.50 g/l and higher at predominance of crystals as suppressed dendrites upon a crystallographic picture one should diagnose a malignant cerebral tumor, and at protein concentration being below 0.50 g/l and predominance of crystals as branched dendrites - a benign cerebral tumor. The innovation enables to fulfill not only due diagnostics of cerebral neoplasms, but detect the degree of tumor process malignancy at the early stages of its development.
 Method for investigation of histological specimen surface relief / 2270446
Claimed method includes surface relief investigation by using plane field microscope in reflected light at one-side falling shadow-forming lighting. Light-reflecting ability of specimen surface is provided by silver impregnation. Said impregnation is carried out by specimen hold in 10 % silver nitrogen solution for 10 min followed by reducing thereof with 1 % ascorbic acid solution for 1 min.
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FIELD: biotechnologies. SUBSTANCE: identification method of blue pus bacillus Pseudomonas aeruginosa involves inoculation of the test material on a hard substrate, incubation of the inoculum under anaerobic conditions at the temperature of 37°C during 16-18 hours. The obtained bacterial mass in the amount of one bacteriological loop is placed in 300 mcl of a physiological solution and warmed-up at the temperature of 98-99°C during 20-30 minutes and centrifuged at 12000 revolutions per minute during 30 seconds. To a supernatant there added is colouring material for electrophoretic detection in the quantity of 0.5 mcl and 20 mcl is added to a well with a size of 4×1 mm 1.2% agarose gel on TAE-buffer with 10 mcl of 1% ethidium bromide. Besides, the amount of 3 mcl of the solution of standard DNA-marker 1 kb containing DNA fragments in the range of 250-10000 bp is added to the test well. Horizontal electrophoresis is performed during 15-20 minutes, and at detection on the obtained electrophoregramme of the test bacterial mass of three strips radiating in ultraviolet light, one of which corresponds to fragments of standard DNA-marker with the size of 10000 bp, the second one corresponds to fragments of standard DNA-marker with the size of 6000-8000 bp and the third strip at the end of the track in the form of a whisk, which corresponds to fragments of standard DNA-marker with the size of less than 750 bp, there identified is Pseudomonas aeruginosa in the test bacterial mass. EFFECT: method allows quick and complete identification of pigment-shaping and non-pigment strains Pseudomonas aeruginosa in a test bacterial mass. 5 dwg, 2 ex
The invention relates to medicine, namely to molecular biology, Microbiology and surgery, and can be used for diagnostic purposes to identify Pseudomonas aeruginosa (Pseudomonas aeruginosa) in clinical biomaterial. Wand blue-green pus - Pseudomonas aeruginosa (Ps. aeruginosa is the type species of the genus Pseudomonas. It is widely distributed in the environment. In small quantities it is found in the intestines of humans and animals, are found in the skin, mucous membranes, various household items, soil, water. Ps. aeruginosa can cause a variety of suppurative diseases, especially in patients with low resistance to infection: suppuration of wounds, osteomyelitis, otitis, pneumonia, pulmonary abscesses, as well as food poisoning, endocarditis, sepsis, and other Frequently Ps. aeruginosa can be found in nonhealing, bad generousity wounds, trophic ulcers, bedsores. Ps. aeruginosa is one of the most frequent causative agents of infections associated with health care, which are characterized by severe and associated with high mortality. High biochemical activity and a wide range of enzymes provide Pseudomonas aeruginosa adaptation to diverse conditions of existence in the environment. Polanti-beatekostenlose, the ability is long with manatsa in the environment, including when exposed to different chemicals, etc. that characterize this microorganism as an ideal hospital pathogen. In most cases, to diagnose lesions of the internal organs by Escherichia coli requires the allocation of the pathogen. The main method of laboratory diagnosis of Ps. aeruginosa - bacteriological examination with subsequent identification, which take into account the ability of a microbe to the formation of pigments and biochemical properties. General principles the identification of Ps. aeruginosa include [Manual of Clinical Microbiology 7th ed., ed. in chief PR. Muttay. Washington: Am. Soc. Environ., 1999; R.]: - application environments General purpose and differential diagnostic media for the initial allocation of bacteria and prior to their differentiation from fermenting glucose gram-negative bacteria; - study of the morphology of cells per gram stain; - screening of the cardinal signs of a selected culture - production of the pigment, the presence of mobility, cytochrome oxidases, sacharolytica activity on oxidative-fermentative (OF) environment Hugh Leifson with glucose; - the use of additional biochemical and other tests for identification. The disadvantages of this method are: 1) the frequency of correct identification of these microorganisms in clinical laboratories does not exceed 30-60%, because the diagnostic is rsnake non-fermentative bacteria, including Ps. aeruginosa, volatile and unstable; 2) schema research mnogoetapnoe, bulky, large number of tests are difficult to perform; 3) significantly complicates the isolation and identification of Pseudomonas aeruginosa that it usually is in the studied material not in monoculture and in Association with other microorganisms, most often with Proteus, which increases the duration of laboratory research; 4) since the formation of pigments Ps. aeruginosa is considered by a number of authors [Ruban E. L. Physiology and biochemistry of the genus Pseudomonas. - M.: Nauka, 1986. - 200 S.; Belyakov C. D., Rapes L. A., Ilyukhin Century. And. Found and Pseudomonas. - M.: Medicine, 1990, 224 S.] as the key and a dominant feature at vnutrirodovoe differentiation, the difficulties in the diagnosis of Ps. aeruginosa associated with a large number of its non-pigmented strains. So, in the early 60-ies of the share of non-pigmented strains of Pseudomonas aeruginosa accounted for 18-25%, and by the early 90-ies is already 50%. Non-pigmented Pseudomonas aeruginosa can be difficult to distinguish from other non-fermentative bacteria belonging to the genera Acinetobacter, Moraxella, Flavobacterium. To identify non-pigmented strains of Pseudomonas aeruginosa requires the production of additional tests: variousfields, cytochrome oxidase, nitroreductase, gelatinase, argini is dihydrolase, isindeterminate, the oxidation of glucose and mannitol in the medium Hugh Leifson, induction of production of pyocyanin on the environment king and pyoverdine on the environment king Century [Zubkov, M. N. Non-fermentative bacteria: classification, General characteristics, role in human pathology. Identification of Pseudomonas spp.and similar micro-organisms // Infection and antimicrobial therapy. - 2003. - So 5. No. 1. - S. 24-30; the Ministry of health of the USSR №535 from 22.04.85 "On the unification of microbiologic methods used in clinical diagnostic laboratories medical institutions". - S. 109-115]. However, each test has limitations, as it provides an opportunity to identify only those strains that have these properties. The well-known scheme of highly specific bacteriological identification and differentiation of atypical and non-pigmented strains of Ps. aeruginosa from objects in the external environment, food raw materials and food products with cumulative medium with succinate sodium, arginine and furadonin and improved selective medium with cetrimide and selected tests bacteriological identification, which is carried out for 75 hours, with further typing by using specific bacteriophage for 19-20 hours (Shestakov, A., Improvement of methods for the isolation, identification and indie the emission of the bacteria Pseudomonas aeruginosa. The dissertation on competition of a scientific degree of candidate of biological Sciences, 2010 - 22 pages). The disadvantage of this method is the complexity of the technological process complexity, large duration of infusion, using a large variety of reagents. A known method for the diagnosis of nosocomial strain of Pseudomonas aeruginosa Ps. aeruginosa, which includes the determination of the sensitivity of the strain to antibiotics and its pageType. Additionally determine the stability of the strains to disinfectants, its plasmid profile, serotype, adhesion to epithelial cells and to be diagnosed with a strain of Pseudomonas aeruginosa Ps. aeruginosa as a hospital in the absence of sensitivity to the strain of not less than 9 antibiotics, the same fago - and serotype, resistance to at least five disinfectants, similar plasmid profile and the coefficient of adhesion to epithelial cells 15±0.2 and more (Shkarin centuries, Nikiforov, C. A., Vorobyov O. N., Davydova N. A., Sargin is protected by E. C. method for the diagnosis of nosocomial strain of Pseudomonas aeruginosa Ps. aeruginosa. The application for the invention 94026116/13, publ. 20.08.1996). The disadvantage of this method is the complexity of the technological process. Thus, despite the fact that the demand in the diagnosis of Pseudomonas aeruginosa infection is obvious, existing bacteriological parameters of the schemes, methods of separation of the Oia, identify and display Ps. aeruginosa imperfect. Closest to the claimed method is a method of allocating non-pigmented strains of Pseudomonas aeruginosa (Nadtoka C. L., volians Y. L., Vorobyov O. N., Bozhko N. G., Grishko A. A., Kirichenko, I. A., Redko, I. A., Gogolev I. C., N. Ryazanov.In., Tooth L. I., V. E. A. Copyright certificate №1442550, publ. 07.12.88,) by planting researched material on nutrient medium, followed by incubation of the culture and the study of growing crops for the presence of pigment. To speed up the way the seeding implement mastopathy agar and/or broth, which is directly before sowing add chlorophyllipt a dose of 12.5-25 µg/ml of medium, and the incubation carried out for 16-18 hours At the upper layer of the medium is colored in blue-green color inherent in Pseudomonas aeruginosa. The authors believe that their method allows you to select non-pigmented strains of Pseudomonas aeruginosa in 2 times faster and 6 times more effective compared with the medium A. king However, the environment king And used to enhance the ability of Pseudomonas aeruginosa to produce only two pigment - blue-green pigment pyocyanin and in some cases porubin giving red staining. Authors ' method is also determined by only the strains generated at the production only of pyocyanine, and the strains producing other is their pigments - the pyoverdine (brown color), porubin (bright red), fluorescein (fluorescent UV-rays), this method is not defined. The present invention is to develop a laboratory method for the identification of Pseudomonas aeruginosa, which will more fully and accurately diagnose and identify how pigmented and non-pigmented strains of Ps. aeruginosa due to the detection of bacterial DNA as an essential structural and functional component of the cell. This object is achieved by a method for the identification of Pseudomonas aeruginosa Pseudomonas aeruginosa, including planting researched material on a solid nutrient medium, incubation planting in aerobic conditions at 37°C for 16-18 hours. The obtained bacterial mass in the number of single loops are placed in 300 μl of saline solution and heated at 98-99°C for 20-30 minutes, centrifuged at 12000 rpm for 30 seconds. To the supernatant add dye for electrophoretic detection in the amount of 0.5 µl and 20 µl make the hole the size of 4×1 mm to 1.2% agarose gel in TAE buffer with 10 ál of 1% ethidium bromide. In parallel in the control hole making 3 μl of solution of standard DNA marker 1 kb containing the DNA fragments in the range 250-10000 bp. Spend a horizontal electrophoresis for 15-20 minutes, and when detecting the received electrophoregram studied bacterial mass three glowing in ultraviolet light strips, one of which corresponds to the fragments of the standard DNA marker 10,000 bp, the second corresponds to the fragments of the standard DNA marker size 6000-8000 bp and the third strip at the end of the track in the form of "panicles", corresponding to the fragments of the standard DNA marker size of less than 750 bp, identify Pseudomonas aeruginosa in the study of bacterial mass. The novelty of inventions - Received the bacterial mass in the number of single loops are placed in 300 μl of saline solution and heated at 98-99°C for 20-30 minutes, centrifuged at 12000 rpm for 30 seconds. To the supernatant add dye for electrophoretic detection in the amount of 0.5 µl and 20 µl make the hole the size of 4×1 mm to 1.2% agarose gel in TAE buffer with 10 ál of 1% ethidium bromide. In parallel in the control hole making 3 μl of solution of standard DNA marker 1 kb containing the DNA fragments in the range 250-10000 bp. - Spend a horizontal electrophoresis for 15-20 minutes, and when detecting a received electrophoregram studied bacterial mass three glowing in ultraviolet light strips, one of which corresponds to the fragments of the standard DNA marker 10,000 bp, the second corresponds to the fragments of the standard DNA marker size 6000-8000 bp and the third strip at the end of the track in the form of "PI", sootvetstvujushej fragments of standard DNA marker size of less than 750 bp, identify Pseudomonas aeruginosa in the study of bacterial mass. In the patent and scientific literature there is no information about a similar method for the identification of Ps. aeruginosa. The essential features of the invention allows to obtain a new technical result consists in the rapid and complete identification pigmentocracy and non-pigmented strains of Pseudomonas aeruginosa in the studied bacterial mass, obtained by cultivation on solid nutrient media. The method allows to differentiate Pseudomonas aeruginosa from other representatives of non-fermentative gram-negative bacteria (Acinetobacter spp.) and the most important genera and species of the family Enterobacteriaceae. The invention is illustrated in the photographs of Fig.1 - Fig.5, which presents the results of identification of Ps. aeruginosa in bacterial mass grown standard from various sources of the biomaterial of the proposed method. In Fig.1 presents a photograph of electrophoregram DNA sample Pseudomonas aeruginosa identified by the proposed method, after electrophoresis in 2% agarose gel. In Fig.2 presents a photograph of electrophoregram samples of cultures of different genera or strains of bacteria, including Ps. aeruginosa, processed by the proposed method, after electrophoresis in 1.2% agarose gel: Klebsiella spp. No. 11, Enterobacter spp. No. 23, Enterobacter spp. No. 22, Acinetobacter baumannii No. 18,Acinetobacter baumannii No. 10, Acinetobacter baumannii No. 51, Ps. aeruginosa No. 42 non-pigmented strain, Ps. aeruginosa No. 3, Ps. aeruginosa No. 2, Ps. aeruginosa No. 1 non-pigmented strain, Escherichia coli No. 5, Escherichia coli No. 6, Escherichia coli lactosome-negative No. 7, Escherichia coli No. 4, Proteus mirabilis No. 17., Proteus mirabilis No. 16. In Fig.3 presents a photograph of electrophoregram DNA sample Ps. aeruginosa, selected by the proposed method in comparison with standard DNA marker, and a figure showing the structure of a standard DNA marker (OOO "Laboratory MEDIGEN", , Novosibirsk). In Fig.4 presents a photograph of electrophoregram DNA samples of different strains of Ps. aeruginosa compared with DNA samples Proteus mirabilis and Escherichia coli isolated from a wound of a patient burn units. In Fig.5 presents a photograph of electrophoregram the DNA of the bacteria isolated from lymphedema of the right lower limb II senior patient surgery. Holes for introducing DNA samples, tracks samples DNA - B, the DNA band corresponding to the DNA fragments marker 10,000 bp, the DNA band corresponding to the DNA fragments marker size 6000-8000 bp - b, band DNA in the form of "panicles" corresponding to the DNA fragments as a marker of less than 750 bp - century During the research we discovered that from scraping grown on nutrient agar culture of Pseudomonas aeruginosa, placed in the number 1 BAC is biologicheskii loop in 300 μl of saline solution and heated at 99°C for 30 minutes, outstanding DNA, after which horizontal electrophoresis in 2% agarose with bromide by ethidium bright aluminium under ultraviolet light with the formation of three fluorescent zones (bands) - next to the hole (a), near the hole (b) and at the end of the track in the form of "PI" () (Fig.1). The comparative analysis of DNA samples of various genera and strains of bacteria that were isolated and analyzed by the proposed method, but using a 1.2% agarose electrophoresis. In Fig.2 presents a photograph of electrophoregram DNA samples following strains of bacteria - Klebsiella spp., No. 11 (lane 1); Enterobacter spp., No. 23 (lane 2); Enterobacter spp., No. 22 (lane 3); Acinetobacter baumannii No. 18 (lane 4); Acinetobacter baumannii No. 10 (lane 5); Acinetobacter baumannii No. 51 (lane 6); Ps. aeruginosa No. 42 non-pigmented strain (lane 7); Ps. aeruginosa No. 3 (lane 8); Ps. aeruginosa No. 2 (track 9); Ps. aeruginosa No. 1 non-pigmented strain (lane 10); Escherichia coli No. 5 (track 11); Escherichia coli No. 6 (track 12); Escherichia coli of lactosuria-tive No. 7 (track 13); Escherichia coli No. 4 (track 14); Proteus mirabilis No. 17 (track 15); Proteus mirabilis No. 16 (track 16). This electrophoregram shows that on the tracks with samples of DNA of different strains of Ps. aeruginosa as pigmentocracy (track 8, 9) and non-pigmented (track 7, 10) strains, clearly reveals the same glow as three bands (a, b, C), while on the tracks with samples of other genera and species of bacteria so the glow is missing. This allowed to conclude that in contrast to other studied species of bacteria in this experiment, from the culture of Ps. aeruginosa proposed method are DNA molecules of different lengths, after which horizontal electrophoresis easily identified under ultraviolet light and give a typical picture of the glow. Using a 1.2% agarose for electrophoresis (instead of 2%) led to an increase in the distance between the strips in 15-minute electrophoresis and obtaining thus a clearer picture of the glow, the characteristic entered in the formula. To estimate the size of DNA molecules of Ps. aeruginosa detected by the proposed method and visible on electrophoregram as separate glowing in ultraviolet light strips, was the standard used to determine the length of double-stranded DNA molecules - DNA marker 1 kb, consisting of 13 DNA fragments in the size range 250-10000 bp, the calibration of which is shown in Fig.3, lane 3. (Manufacturer DNA marker LLC "Laboratory of MEDIGEN", , Novosibirsk). When comparing the received electrophoregram sample of this DNA marker (Fig.3, lane 1) and DNA sample Ps. aeruginosa (Fig.3, lane 2), selected by the proposed method with standard calibrated sizes of the DNA molecules (Fig.3, lane 3) was that linked to electrophoregram three stripes DNA Ps. aeruginosa (a, b, is; track 2) correspond to fragments of the standard DNA marker sizes 10000 bp, 6000-8000 bp and less than 750 bp (Fig.3, lanes 1 and 3), the characteristic introduced us to the formula. Thus, the proposed method allows for 1 hour to identify as pigmentbased and non-pigmented strains of Ps. aeruginosa grown on solid nutrient medium for 16-18 hours, and to differentiate them from other representatives of non-fermentative gram-negative bacteria (Acinetobacter spp.) and the most important genera and species of the family Enterobacteriaceae. The method is as follows. The fence and the planting of clinical material spend the conventional methods. The obtained bacterial culture taken from the surface of a simple solid nutrient medium (mastopathy agar, Wednesday Endo) bacteriological loop (single loop) and placed in a disposable plastic container with cover and lock containing 300 μl of saline. The microbial suspension is shaken (on the vortex for 10-15 seconds) and is heated in the solid-state thermostat or a water bath for 20-30 minutes at 98-99°C. If testing bacterial mass is postponed indefinitely, tubes, frozen at -20°C until use. Further contents of the test tubes thawed, shaken and heated, as described above. After prograni the suspension is centrifuged at 12000 rpm for 30 seconds, add dye for electrophoretic detection (NGOs "Litex") in the amount of 0.5 µl per sample. Take about 20 μl of the supernatant, which is poured into a hole size of 4 mm × 1 mm to 1.2% agarose gel prepared in 20 ml of TAE buffer with pre-added to the gel 10 ál of 1% ethidium bromide. To assess the structure of electrophoregram DNA using the standard to determine the length of double-stranded DNA molecules in the range 250-10000 bp used during electrophoresis in agarose gels, DNA marker 1 kb, consisting of 13 DNA fragments in the range 250-10000 bp (OOO "Laboratory MEDIGEN", , Novosibirsk). Standard DNA in the amount of 3 μl poured into a separate well of the gel. Spend a horizontal electrophoresis for 15-20 minutes, controlling the duration of visual motion streaks of dye, which must pass from one hole 1.5 cm After electrophoresis the gel is placed on the glass of the UV-transilluminator and analyze the result in passing ultraviolet light with a wavelength of 310 nm. On electrophoregram detect the fluorescence of DNA in the form of three horizontal stripes - next to the hole that corresponds to the fragments of DNA marker size 10000 bp, close to the wells, which corresponds to the fragments of DNA marker size 6000-8000 bp and at the end of the track in the form of "PI", which corresponds to the DNA fragments as a marker of less than 750 bp. Three glowing the hair on electrophoregram indicates the presence in the studied bacterial mass Pseudomonas aeruginosa. The absence of luminous allows you to give a negative result. The identification of Ps. aeruginosa in grown on agar bacterial culture takes about 1 hour. Example 1. From the wound of a patient burn Department of the city clinical hospital №1, Novokuznetsk isolated on blood agar and identified by cultural and biochemical tests Proteus mirabilis, Escherichia coli, Escherichia coli lactobacteria and Ps. aeruginosa. 1 loop 18-hour cultures of the causative agents were placed in separate tubes containing 300 μl of saline. All samples were warming at 99°C for 30 minutes, was ofcentrifugal at 12000 rpm for 30 seconds, added dye for electrophoretic detection. 20 µl of supernatant from each tube were poured into the wells of a 4 mm × 1 mm to 1.2% agarose gel prepared in 20 ml of TAE buffer with pre-added to the gel 10 ál of 1% ethidium bromide. After electrophoresis for 20 minutes, the gel was placed on a glass UV transilluminator and analyzed the results. On electrophoregram (Fig.4) it is seen that the luminescence of the samples of DNA of different strains of Ps. aeruginosa (tracks: 1 - amelanotic clinical strain Ps. aeruginosa; 2-5 - clinical strains of Ps. aeruginosa with different pigments), selected by the proposed method, detected in the form of three horizontal stripes near the hole (a), at some distance from the hole (b) and at the end of the tre is a (in). DNA Proteus mirabilis (track 6), Escherichia coli (lane 7), Escherichia coli lactobacteria (track 8) lights similar to DNA Ps. aeruginosa only at the end of the track. Thus, the DNA sample only Ps. aeruginosa, highlighted the proposed method has a characteristic glow in UV, consisting of three bands - a, b and C. Example 2. Patient A., 1961 R., appealed to the surgery complaining of the presence of an ulcer on the right leg after suffering a bullous form of faces. At the time of inspection: lymphedema of the right lower limb II century, a plague, which occupies the front of the semicircle of the right tibia, the dimensions of 20×19 cm, depth up to 1.5 cm, with saped edges, with no signs of reparative regeneration and with abundant determinovanii lymph. At the bottom of the ulcer is tightly fixed white bloom. On the dressing detachable wounds acquires a greenish tint, and has a characteristic sweet smell. Diagnosis: giant lymphedematous trophic ulcers. Conducted fence separated wounds for microbiological testing in the bacteriological laboratory at. Revealed Klebsiella pneumoniae. For specification and verification of the diagnosis was made bacteriological seeding wound patient A. on solid nutrient media: blood agar, yolk-salt agar and environment Endo. During the subsequent standard identification grown colonies selected mixed Kul is ur consisting of strains of Pseudomonas aeruginosa Ps. aeruginosa with lemon pigment and Escherichia coli. At the same time, the bacterial culture grown from a wound patient A. on a solid nutrient medium, in the amount of one bacteriological loop was placed in a test tube with 300 μl of saline solution and subjected to the procedure of heating, ultracentrifugation and electrophoresis in 1.2% agarose, in accordance with the proposed method. Solution of standard DNA marker in the amount of 3 μl were placed in separate wells of the gel. The results are presented on electrophoregram (Fig, 5), on which the following tracks are samples: 1 - DNA marker, 2 - Proteus mirabilis No. 5, 3 - Escherichia coli (sown from the wound); 4 - pigmentocracy strain Ps. aeruginosa (seeded from a wound). It is seen that the DNA on the track 4 aluminium, forming three bands near the hole (and corresponds to fragments of DNA marker 10,000 bp), close to the wells (b, corresponds to fragments of DNA marker size 6000-8000 bp) and at the end of the track (corresponds to fragments of DNA marker size of less than 750 bp). Thus, the identification of Ps. aeruginosa in the material taken from the wound of the patient And, with the help of the proposed method proved to be more reliable in comparison with the conclusion of the clinical bacteriologists. A method for the identification of Pseudomonas aeruginosa Pseudomonas aeruginosa, including planting of the investigated material solid is Yu nutrient medium, incubation of sowing in aerobic conditions at 37°C for 16-18 h, characterized in that the bacterial mass in the number of single loops are placed in 300 μl of saline solution and heated at 98-99°C for 20-30 min, centrifuged at 12000 rpm for 30 s, the supernatant add dye for electrophoretic detection in the amount of 0.5 µl and 20 µl make the hole the size of 4×1 mm to 1.2% agarose gel in TAE buffer with 10 ál of 1% ethidium bromide, in parallel in the control hole making 3 μl of solution of standard DNA marker 1 kb containing the DNA fragments in the range 250-10000 bp, spend a horizontal electrophoresis for 15-20 min, and detection on the received electrophoregram studied bacterial mass three glowing in ultraviolet light strips, one of which corresponds to the fragments of the standard DNA marker 10,000 bp, the second corresponds to the fragments of the standard DNA marker size 6000-8000 bp and the third strip at the end of the track in the form of "panicles", corresponding to the fragments of the standard DNA marker size of less than 750 bp, identify Pseudomonas aeruginosa in the study of bacterial mass.
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