Method of blood plasma proximate analysis for cardiomyoglobin by means of electrochemical immunosensor
SUBSTANCE: method of blood plasma proximate analysis for cardiomyoglobin by means of an electrochemical immunosensor consists in the fact that a blood plasma sample is applied on a surface of a main electrode pre-modified by colloidal gold and monoclonal cardiomyoglobin antibodies; the prepared sample immunosensor is kept, preincubated in a buffer solution; cardiomyoglobin is electrochemically recorded in the sample by the prepared calibration diagram under certain conditions.
EFFECT: more sensitive analysis procedure, and reduced analysis time.
2 dwg, 2 tbl, 4 ex
The invention relates to the field of analytical chemistry, Elektrokhimiya and medical diagnostics and relates to a method of rapid determination of cardiomegaly in plasma using electrochemical immunosensor.
The invention can be used in medical practice for the diagnosis of myocardial infarction (mi). According to statistics, cardiovascular diseases occupy the first place among the causes of death of the population in Russia (www.gks.ru and cause almost half of the total number of deaths in Europe (www.ehnheart.org). Inefficiency or lack of early diagnosis also directly affects the situation. Over the last decade, a large number of studies aimed at the identification of cardiometer, which can be used for diagnosis for heart pain. Cardiometer is a biological substance, raising the level in the blood is observed for cardiovascular diseases or immediately after damage to the heart muscle . Today we know 177 compounds markers of cardiovascular diseases and research in this area continues .
Determination of biomarkers in the blood of patients to diagnose THEM has become a standard procedure in medical practice abroad and is used in leading clinics of the Russian Federation. Nevertheless continuously conducted by the SC of new cardiometer, and a variety of statistical studies confirming the effectiveness of test-systems for diagnostics of THEM. At the present time to diagnose THEM widespread troponin I and T, creatine kinase-MB and cardiomegaly. Serum troponin and creatine kinase-MB point to the death of myocardial cells. Whey cardiomegaly is one of the earliest biomarkers THEM. Cardiomegaly - hemoprotein with a molecular weight of 17,800 and due to its small size, it is rapidly released from damaged tissue in the blood. Its half-life in plasma is only 8.9±1.5 minutes Among patients with acute period of THEM the overall picture of changes in the concentration of cardiomegaly plasma with similar dependencies for creatine kinase-MB and troponin. However, the release of troponin and creatine kinase-MB slower, allowing you to detect them in the blood only after 4-6 hours after onset of symptoms, some patients have increased levels of these markers does not manifest within 12 hours. Whey cardiomegaly appears in the blood within 1 to 4 hours. Serum concentrations of cardiomegaly higher after the damage of various tissues (especially skeletal muscle) or taking cocaine, or in patients with renal failure by reducing clearance (coefficient of purification). However, m is her early racing level cardiomegaly and its ability to determine the acute THEM before other biomarkers compensates for these disadvantages, especially in combination with troponin and creatine kinase-MB, and conducting a series of tests [3, 4].
In various sources, the threshold concentration of cardiomegaly in the blood, the excess of which proves THEY vary in a fairly wide range (from 70-90 µg/l [5-8] to 200 µg/l ) and seems to depend on the individual characteristics of the organism. However, the average threshold of 100 µg/l adhere to most researchers [10-14].
In today's market presents the following tests and system table immunoassay proteins cardiometer produced by foreign manufacturers: Stratus® CS STAT (Dade Behring Inc), (i-STAT® (Abbott), Triage® Cardiac Panel (Biosite), RAMP® (Response Biomedical Corp), Cardiac Reader™ (Roche), PATHFAST® (Mitsubishi Chemical Europe GmbH), MGL-CHECK-1 (Vedalab), HEXAGON TROPONIN (HUMAN GmbH).
Published work on the definition of cardiomegaly is "sandwichy" immunoassay. The method is based on the selective binding of proteins markers corresponding antibodies attached to the surface of the sensor. The detection signal is due to the second top layer of antibodies with different labels (PL. 1).
However, all these methods are quite complex, requiring large expenditures of time. None of them are allowed today to clinical application. As can be seen, and the presented data, only one method of immunoassay adapted for use with blood. None of the authors gives no information about working with real samples of patients with THEM. Accuracy and other analytical characteristics definition in this case will depend on many factors: the activity of antibodies, enzyme-labels, substrate concentration, etc.
The redox activity of iron heme serves as a kind of token hemoproteins and allows for their direct detection, and outside influence on the active center of the enzyme. Myoglobin belongs to the family of hemoproteins - electroactive proteins, the molecules of which contain iron ion. The effect of direct electron transfer from the electrode to the heme iron is widely used for manufacturing electrochemical sensors for hydrogen peroxide and nitrite ion. For registration and modification of the electrode surface are used surfactants [19-21], polymer hydrogels , membranophone complexes of polyelectrolytes and other
The aim of the present invention is to develop a method for determining cardiomyogenesis in plasma samples using electrochemical immunosensor. The method of obtaining electrochemical immunosensor for the direct determination of cardiomegaly described in the patent . Previously it was shown that the selective determination of cardiomyo is of Aubin in buffer solutions. The Express-analysis of cardiomegaly in plasma samples required significant revision of the methodology for determining and making immunosensors. The main objectives were: 1) increasing the detection sensitivity and 2) a significant reduction of analysis time.
Unlike most foreign researchers, implementing "sandwich" the analysis scheme, the proposed method is based on own electroactivity of cardiomegaly and direct detection of specific interaction between molecules of hemoprotein and corresponding antibodies. The proposed scheme direct registration of myoglobin due to direct electron transfer from the electrode to the heme iron ion catalyzed oxygen:
The contribution to the signal amplification contribute:
1) gold nanoparticles;
2) the stabilizer of gold nanoparticles - DDAB, absorbing oxygen from the background electrolyte;
3) method of reception signal is a square-wave voltammetry.
The need for the use of antibodies based on the task of selective determination of cardiotomy enzyme released into the blood when the heart muscle is damaged. The analytical signal is the magnitude of the detected current of the electron transfer from the heme iron to the surface of the graphite is of the first electrode, modified by gold nanoparticles. Deposited on the surface of the electrode colloidal gold particles work as an ensemble of microelectrodes and allow not only to increase the area of the active surface of the sensor, but also to improve the ratio signal/noise. The transition from the flat electrode to the system of microelectrodes with radii comparable with the thickness of the diffusion layer, leads to edge effect when the value of the limiting current is directly proportional to the radius of microelectrode.
EXAMPLE 1. The method of preparing a colloidal solution of gold
Preparation of the solution zolotoporfirovoe acid.
Weigh (3.9) mg trihydrate zolotoporfirovoe acid (HAuCl4×3H2(O)transfer sample in Eppendorf and dissolved in 1 ml of distilled water. The solution is stored in Eppendorf at 4°C not longer than 1 month.
Preparation of the solution dimethyldioctadecylammonium bromide (DDAB) (0.1 M).
Weigh (46.3) mg dimethyldioctadecylammonium bromide, transferred to the sample in Eppendorf and dissolved in 1 ml of chloroform. The solution is stored in Eppendorf at 4°C not longer than 1 month.
Preparation of a solution of sodium borohydride (0.4 M).
Weigh (15.2) mg sodium borohydride, carry it in Eppendorf and dissolved in 1 ml of distilled water. The solution is not stored.
Preparation colloides the solution of gold (5 mm).
A round bottom flask is fixed on a tripod over a magnetic stirrer. Into the flask using a pipette, transfer 1 ml of 0.1 M solution DAB. With vigorous stirring to a solution of DAB add 0.5 ml of 10 mm solution zolotoporfirovoe acid, selected by the eyedropper. Then, with vigorous stirring, added dropwise 0.2 ml of freshly prepared solution of sodium borohydride. After 2 hours dyed organic layer is separated from the water.
EXAMPLE 2. Fabrication of electrochemical immunosensors on cardiomyogenic in plasma.
Preparation of 6 M NaOH
Weigh (24.0 g of sodium hydroxide, transfer the sample into a measuring flask of 100 ml and adjusted with distilled water to the mark. Solution store in plastic dishes under a thrust not exceeding 1 month.
Preparation of phosphate buffer solution
Weigh (1.33 g of potassium dihydrophosphate (KH2PO4and (0,29) g sodium chloride (NaCl). Sample salts transferred into a volumetric flask of 100 ml and adjusted with distilled water to the mark. The solution is thoroughly mixed on a magnetic stirrer until complete dissolution of the salts. On the pH meter using 6 M NaOH solution was adjusted pH to 6.5. The solution is stored in a plastic container with the lid tightly closed at 4°C not longer than 1 month.
Preparation of the solution of antibodies to cardiomegaly
The original solution of antibodies to cardiomyo is the Aubin (firm USBio, USA, cat. No. m-16A) were diluted 10 times with phosphate buffer solution pH 6.5±0,1. In a test tube "Eppendorf" transfer an aliquot of 10 µl of the antibody solution and add 90 ál of buffer solution. The solution is stored in a test tube "Eppendorf" at -20±2°C not more than 1 month.
Preparation of electrochemical biosensors for cardiomegaly
On the surface of the graphite working electrode (manufactured using carbon paste firm "Gwent", England; manufacturer firm "Rosens", Moscow) put 1 ál of freshly prepared colloidal solution of gold (5 mm). We allow the chloroform to evaporate for 10 minutes Then put 1 μl of the antibody solution, give the drop to dry for 10 min and incubated for biosensor in the refrigerator with a temperature of 4±2°C during the night. Biosensors stored at 4±2°C not more than 1 month.
EXAMPLE 3. Measuring the concentration of cardiomegaly using electrochemical immunosensor in plasma samples.
Preparation for measurement
On the working surface electrochemical biosensor with a pipette and put a 1 μl sample of blood plasma and placed in an oven with a temperature of 37±1°C in 15 minutes
The parameters of the measurement signal
In the program to the device sets the following parameters of the measurement signal (figure 1):
Method: Quadrantanopia voltammetry;
Incubation time:900 s;
Frequency: 10 Hz;
The initial potential: 0.1;
The final potential: -0.6 V;
Step potential: 0.005;
The signal measurement
In the cell make 1 ml of phosphate buffer solution. Fix biosensor coated with a break in the measuring cell, immersed in a buffer solution and begin the procedure of the measurement signal. The obtained voltammetric curve remembered as a separate file.
The calculation of the concentration of myoglobin
If the received voltamperometry peak is observed (Emax=-200...-300 mV), then using the device to produce a definition of the area received the maximum recovery of myoglobin. On the previously constructed calibration curve to determine the concentration of myoglobin, corresponding to the received value of the peak area.
EXAMPLE 4. The definition of cardiomegaly in plasma samples of patients with myocardial infarction and healthy donors.
The developed biosensor and method for rapid determination of cardiomegaly were tested on 5 plasma samples of patients and healthy people. Table 2 shows the concentrations of cardiomegaly obtained immediately after blood sampling using the RAMP system (Response Biomedical Corp), for each of the samples according to the instructions to the device. The obtained samples were stored at -70±2°C and were thawed C is directly before measurements.
|Characteristics of plasma samples|
|ID sample||The concentration of cardiomegaly, ng/ml|
The following figure 1 and 2 shows a typical square-wave voltamperometry reduction of iron of heme cardiomegaly obtained according to the developed method of Express-analysis for different samples: serum without cardiomegaly (control sample), serum from 14 µM by kardiomiogeneza and plasma sample of the patient NAMED (ID 0001) (Fig 1)and also shows a linear relationship between the concentration of cardiomegaly determined using the analyzer RAMP® (Response Biomedical Corp, Canada) and signal immunosensor (peak area reduction of iron of heme cardiomegaly)obtained according to the developed methodology (figure 2).
The method of rapid determination of cardiomegaly in plasma using electrochemical immunosensor, namely, that on the surface of the working electrode, pre-modified colloidal gold and monoclonal antibodies to cardiomegaly, put a 1 μl sample of blood plasma, can withstand the resulting immunosensor with a break of 15 min at 37±1°C, perform incubation for 15 min in the working phosphate buffer solution at rn,5±0,1, conduct electrochemical check cardiomegaly by measuring the peak area recovery heme iron by the method of square-wave voltammetry and determine the content of cardiomegaly in the sample prior calibration.
SUBSTANCE: method of detecting anthrax pathogen in environment and biological fluids is based on functional detection of activity of one of anthrax toxin components, weak protease of anthrax lethal factor (LF). Applied principle of highly sensitive detection of LF proteolytic activity is based on the system of amplification of sugnal, originating from the act of substrate cutting, by means of present in substrate composition auxiliary enzyme of alkaline phosphatase of Escherichia coli (AP). In composition of recombinant LF substrate in addition to alkaline phosphotase and specific for LF substrate sequence RRKKVYPYPME, there is a peptide which is biotinilated in vivo and in vitro by means of biotin-ligase of E.coli and ensuring substrate immobilisation on solid phase (plaque surface) due to interaction with avidin and its derivatives, and sequence of six histidines for purification of recombinant substrate from periplasm of E.coli by means of metal-chelate resin.
EFFECT: method in accordance with the invention possesses high sensitivity, with application of fluorescein diphosphate makes it possible to detect up to 1 pM of lethal factor, rapidity of execution, high specificity, low risk of false-positive signals.
1 dwg, 3 ex
SUBSTANCE: set of reagents for quantitative determination of avermectins contains an ivermectin conjugate with bovine serum albumin which is immobilised on a polystyrene dish, a peroxidase conjugate of highly specific mouse monoclonal antibodies to ivermectin, and ivermectin calibration samples (with concentration between 0 and 100 ng/ml). The monoclonal antibodies in the set are produced by a strain of hybrid cultured cells of Mus. Museums L., which are deposited in the Collection of transferred vertebrate somatic cells of the D. I. Ivanovsky Research Institute of Virology under No. 05/09. From non-specific components, the set contains a buffer for culturing the monoclonal antibody conjugate with horseradish peroxidase, a buffer for culturing the analysed samples and washing the trays, reagents for detecting peroxidase activity, a substrate solution, hydrogen peroxide and tetramethylbenzidine and a stop solution (1M sulphuric acid). The disclosed set is designed to detect residual quantities of avermectins in tissue and biological fluids for monitoring chemical contamination of animal products.
EFFECT: use of the set enables to determine main components of an avermectin complex in animal products, while providing high sensitivity and specificity of detection.
1 dwg, 2 tbl, 4 ex
SUBSTANCE: proposed device comprises sensitive surface consisting of binding sections that allow specific attachment to biological objects with magnetic labels, magnetic pickup element, appliance to differentiate between magnetic labels connected to binding sections and magnetic labels, not bound, to act on binding sections. System to determine concentration of target objects consists of measuring device and electronic circuit to detect changes in magneto resistive resonance of magnetic pickup element. Note here that electronic circuit stays in or out of substrate. To determine concentration of target objects in fluid, fluid medium containing magnetic labels is subjected to magnetic field to differentiate between bound and unbound magnetic labels for period when binding process occurs on sensitive surface.
EFFECT: higher accuracy and rate of determining concentration of target molecules in fluid.
24 cl, 9 dwg
SUBSTANCE: method of obtaining erythrocyte diagnosticum for reaction of indirect hemagglutination (RIHG) in case of cattle coccielosis consists of fractional formalinisation of sheep erythrocytes and their sensibilisation with coccielosis antigen at 70°C for 30 minutes. For sensibilisation used are erythrocytes, loaded with sensitin obtained from vaccine strain C.burnetii M-44, heated on water bath at 70°C for 30 minutes, with further triple washing of erythrocytic diagnosticum with buffer solution with pH -7.2.
EFFECT: invention ensures increase of specificity and activity of diagnosticum in RIHG in case of cattle coccielosis.
SUBSTANCE: claimed is method of obtaining reactive sugar which includes the following stages: i) providing sample, containing reducing sugar; ii) providing solid substrate covalently bound to linker, which includes catch group containing -NH2 group, in which said linker is optionally bound to said solid substrate via spacer; iii) reaction of said reducing sugar with said -NH2 group resulting in obtaining immobilised sugar; iv) reaction of free -NH2 groups with camping-agent, with camping-agent containing reactive group able to react with -NH2 group and v) reduction of C=N bonds with reducer resulting in obtaining reactive sugar of SugarCHn-NH- structure, bound with solid substrate via linker and, optionally, spacer, where n is 1 or 2 manipulation with immobilised carbohydrates by derivation.
EFFECT: improvement of method
45 cl, 2 tbl, 7 ex, 25 dwg
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention concerns development of a diagnostic test system in immunochip format and a method of simultaneous and differential detection of reaginic antibodies and antibody spectrum to diagnostically significant mmunologically relevant proteins Treponema pallidum of G (IgG) and M (IgM) classes. The diagnostic test system in immunochip format for differential serum diagnostics of syphilis consists of an immunosorbent with separately immobilised antigens Treponema pallidum Tp15, Tp17, TmpA, Tp47, conjugate and reactants required to detect an antigen-antibody complex. Cardiolipin is additionally immobilised on the immunosorbent; antigens Treponema pallidum and cardiolipin are immobilised at least, in two repetitions, and conjugate consists of mixed antispecies human IgG antibodies and human IgM antibodies modified by two phosphors with different spectral characteristics. On the immunosorbent, there can be additionally immobilised at least, one antigen and/or peptide Treponema pallidum specified e.g. of Tp39, Tp41, Tp42, Tp44.5, Tp92, Tp0453 at least in two repetitions. Cardiolipin can represent an oxidised cardiolipin derivative bounded with protein. The differential serum syphilis diagnostic technique with using the diagnostic test system in immunochip format involves application of human IgM modified by two phosphors with different spectral characteristics. On the immunosorbent, there can be additionally immobilised at least one antigen and/or peptide Treponema pallidum, specified e.g. of Tp39, Tp41, Tp42, Tp44.5, Tp92, Tp0453 at least in two repetitions. Cardiolipin can represent an oxidised cardiolipin derivative bounded with protein. The differential serum syphilis diagnostic technique with using the diagnostic test system in immunochip format implying that a cultivation solution for check and test samples is introduced on the immunosorbent with separately immobilised antigens Treponema pallidum and cardiolipin; the check and test samples are introduced; the prepared mixture is incubated at temperature 20-42°C for 15-60 min to prepare the antigen-antibody complex; the immunosorbent is rinsed; then a conjugate solution of mixed human IgG antibodies and human IgM antibodies modified by two phosphors with different spectral characteristics is introduced on the immunosorbent; it is followed with incubation at temperature 20-42°C for 15-60 min; the immunosorbent is rinsed, dried to detect the prepared antigen-antibody complex.
EFFECT: improved diagnostic accuracy.
20 cl, 5 dwg, 2 ex
FIELD: chemistry; biochemistry.
SUBSTANCE: method for combined immunobiological analysis of cells using a biochip involves incubation of the biochip which contains immobilised antibodies, with suspension of cells, washing the biochip from non-bonded cells, determination of coexpression of antigens on the bonded cells. The obtained result is assessed by determining presence of bonded cells in the region of the stain of the biochip and bonding density of cells and interpretation of the obtained result. Coexpression of antigens on cells bonded to the biochip is determined by carrying out one or more immunocytochemical reactions. When reading out the result, morphological analysis of cells bonded to the biochip is also carried out and presence and character of colouring of cells and their components with the reaction product are determined.
EFFECT: use of the disclosed method provides high reliability and information content of analysis.
9 cl, 6 dwg, 2 ex
SUBSTANCE: after antigen-antibody complex is prepared in a reaction compartment wherein said antigen to be modified and antibody are bound, said reaction compartment is washed with using a sample solution. The invention allows detecting a signal reflecting quantity of said antigen to be modified at precision comparable when using a washing solution to this without using said solution. Herewith, the washing solution is not required to be supplied from the outside of a chip or to be ensured therein beforehand.
EFFECT: immunoassay is easily realised on the chip.
6 cl, 6 dwg, 1 tbl, 2 ex
SUBSTANCE: invention can be used for evaluating the antibody level in blood serum by enzyme-linked immunosorbent assay (ELISA) in diagnostics of the diseases caused by capsular forms of such agents, as Meningococcus, Pneumococcus, Streptococcus, Haemophilus, Neisseria, Salmonella, Klebsiella, Pseudomonas, and also for evaluating the postvaccinal immunity level.
EFFECT: covalent binding of polysaccharide to the protein pre-immobilised on the tray surface ensures reliable and simple antigen fixation that does not involve effect of foreign chemical groups on the analysis results that leads to simplification of the method for tray sensitisation in the ELISA reliability control.
1 dwg, 1 tbl, 3 ex
FIELD: instrument making.
SUBSTANCE: set of invention relates to devices to determine the level of analysed substances in biological fluids. Proposed system comprises (a) measuring device with housing, logical circuit arranged in said housing, visual display arranged on said housing and measuring system arranged therein. It comprises also (b) cartridge incorporating at least one test strip to analyse lateral flow. Note here that said test strip incorporates (i) lateral flow transport matrix, (ii) zone of analysing specific binding on transport matrix to take up fluid specimen to produce required reaction, and (iii) zone of general chemical analysis on transport matrix to take up fluid specimen to produce required reaction. Note also that sizes of cartridges are selected to allow arranging analysed biological fluid substances in measuring device so that measuring system in above described zones on test strip. Mind that transport matrix first and second segments are made from different materials and bound so that said segments does not allow the product formed on first segment to contribute into reaction on second segment. Proposed invention covers other versions of aforesaid device.
EFFECT: higher efficiency, accuracy and reliability.
162 cl, 27 dwg
SUBSTANCE: invention describes a method of evaluating an immunosuppressive response rate in children with chronic glomerulonephritis characterised by determining the peripheral blood cell kinetics by flow DNA-cytometry; evaluating a cell fraction in a proliferation phases not less than 0.74% enables to consider the immunosuppressive therapy to be prescribed, and observing a decreased cell level in proliferation phases in 2-3 weeks after the beginning of administration at least in 2 times, the therapy is considered to be effective; the absence of positive dynamics requires dosage change or preparation replacement.
EFFECT: higher accuracy and reduced number of research injures, derived clinical laboratory criteria of immunosuppressive prescription and prediction of a therapeutic effect.
4 dwg, 1 tbl, 3 ex
SUBSTANCE: invention describes a method of evaluating the heparin concentration in puerperants underwent acute herpes viral infection in the third trimester, characterised by the fact that a piece of placenta is sampled from the puerperants, homogenate is prepared and processed for glycosaminoglycans recovery. Thereafter, the prepared glycosaminoglycans extracts are separated by gel electrophoresis in a polyacrylamide gel, and a heparin percentage is calculated by optical density sensitometry; a herpes viral infective episode is stated by spectrophotometry and shown by increasing titre of herpes virus antibodies.
EFFECT: higher analysis accuracy.
SUBSTANCE: invention relates to medicine, namely, to immunology and clinic laboratory diagnostics, and can be used to predict course of acute respiratory viral infections (ARVI) in children in the first days of disease and timely administration of immunomodulating medications. For this purpose by means of ELISA immunologic indices of spontaneous and induced interferon-γ in vitro (IFH-γ) are determined. Index of interferon-γ stimulation (IS IFH-γ) is calculated by division of index of induced level by index of spontaneous level of interferon-γ. Also carried out is calculation of lymphocyte activation index (LAI IFH-γ) per 1000 lymphocytes by division of index of induced interferon-γ by absolute number of patient's lymphocytes. Additionally in blood plasma determined is content of interleukin-10 (IL-10). If values of IS IFH-γ are higher than 3, LAI IFH-γ equals or is higher than 40, IL-10 is from 30 to 60 pg/ml favourable outcome of disease is predicted with therapeutic treatment which does not include immunomodulators. If IS IFH-γ is lower than 3, LAI IFH-γ is lower than 40, IL-10 is from 60 to 100 pg/ml, predicted are severe course of disease and development of complications, which requires urgent treatment by immunomodelling therapy. If IS IFH-γ is lower than 3, LAI IFH-γ is lower than 30, IL-10 is higher than 100 pg/ml, predicted are severe course of disease with development of bronchopulmonary complications and possible chronisation of pathologic process and recurrent ARVD, which requires additional introduction of immunomodelling medications.
EFFECT: increase of accuracy of early prediction of disease course severity and development of complications in children with ARVI, including those from 1 month old, which makes it possible to carry out necessary anti-viral and immunomodelling therapy, aimed at strengthening immunity cell responses, in due time.
3 tbl, 3 ex
SUBSTANCE: analysed human blood serums are brought in wells of 96-well polystyrene plates with an immobilised complex of pertussis microbe antigens, added with a human IgG3 antibody peroxidase conjugate, added with a substrate mixture that is followed with recording the optical density of the mixture to derive a titre of immunosorbent attached human IgG3 antibodies. The substrate mixture is a ready form of tetramethylbenzidine and hydrogen peroxide. The human antibody titre 1:40 is accepted as pertussis related.
EFFECT: higher sensitivity and specificity of pertussis diagnostics.
1 ex, 2 tbl
SUBSTANCE: not less than twice a week for the first month following the liver transplantation, recipient's peripheral blood is examined for the concentration of circulating stem haemopoetic cells CD 34+ to find its average values for the first 10 and the following 20 days, and if their value is less than 0.1% in the second or both time intervals, a risk of post-transplantation complications is predicted in the recipient.
EFFECT: invention provides high accuracy of prediction of the risk of early, most dangerous post-transplantation liver rejection.
2 ex, 1 dwg
SUBSTANCE: venous blood is taken from a worker exposed to vibration action to analyse a neuron-specific enolase (NSE) level by immune-enzyme assay (IEA). If the NSE concentration is less than 20 ng/ml, the absence of vibration sickness in the person being tested is stated, the NSE level being within 20 to 30 ng/ml shows the presence of degree 1 vibration sickness, while the NSE level exceeding 30 ng/ml indicates degree 2 vibration sickness.
EFFECT: simplified diagnosing of vibration sickness and evaluation of a manifestation degree of vibration sickness.
1 tbl, 3 ex
SUBSTANCE: method of quantitative determination of human chitinase-like protein YKL-40 (another name - human cartilaginous glycoprotein-39, or HcGP-39) is offered. When implementing the method, colloidal chitin or heparin is sorbed in microplate wells, then solutions containing a certain amount of chitinase-like protein, and analysed samples are introduced in the wells. The amount of YKL-40 is analysed by the amount of an enzymatic reaction product formed by interaction of an enzymatic conjugate with YKL-40 antibodies against with a substratum of this enzyme.
EFFECT: method allows well reproducible and high sensitive YKL-40 analysis in various human biopsy materials and excretes.
2 cl, 2 ex, 1 tbl
SUBSTANCE: once admitted to hospital, a patient suffering acute cholecystitis is examined for the blood serum lactoferrin level to calculate a range score; the serum lactoferrin values 200 ng/ml are taken as 1 point; respectively, the lactoferrin level 8 point and lower enable to observe a destructive form of chronic cholecystitis; the lactoferrin level within 8 to 10 points shows catarrhal cholecystitis, and the level 10 points and higher indicates destructive cholecystitis.
EFFECT: use of the technique allows higher diagnostic accuracy in gallbladder tissue destruction in associated acute cholecystitis.
SUBSTANCE: method of determining individual human drug sensitivity or immunotoxicity is based on individual modelling in vitro of the effect on IFN-alpha and/or IFN-gamma production by patient's whole blood leukocytes/lymphocytes. The method involves evaluating a degree of a stimulating/correcting or inhibitory drug action and/or afteraction on the primary IFN system reactivity values.
EFFECT: method provides individual prescription of adequate drugs and thereby higher clinical efficacy for prevention of potential developing medicinal interferon immunodeficiencies.
3 cl, 4 ex
SUBSTANCE: for selective recovery of a viable cell population, a biological fluid sample is placed in a microfluid device cell containing a silicon microchannel matrix with through holes of size 3-30 mcm and channel length 50-300 mcm as a filter medium, and passed through the matrix at rate 0.1-4 ml/min. In case of cell size separation, a cell fraction of the size less than that of the through holes in the matrix, are collected after passing of the microchannel matrix, while the larger cells are eluated from the microchannel matrix channels by eluent backflow. In case of receptor-specific cell separation, a matrix surface is pre-modified by specific cell receptor antibodies, while the target cells are eluated from the matrix channels by isotypic antibodies or haptens.
EFFECT: more efficient and faster selective recovery of the viable cell population from biological fluids.
3 cl, 2 dwg, 4 tbl, 4 ex
FIELD: medicine, ophthalmology.
SUBSTANCE: in lacrimal liquid one should detect the content of interleukin 8 (IL-8) and that of interleukin 1 beta (IL-1β) to calculate prognostic coefficient (PC) due to dividing the first value by the second one by the following formula: At PC value being below 10.0 one should predict favorable disease flow, and at PC value being above 10.0 - unfavorable flow.
EFFECT: higher accuracy of prediction.