Method for determining antibacterial resistance of human body
SUBSTANCE: invention describes a method for determining antibacterial resistance of a human body in diseases caused by a staphylococcus infection, based on blood neutrophil analysis; functional activity of neutrophilic granulocytes of patient's peripheral blood is determined by chemoluminescence with calculating a bacterial activation index (the BAI-index) of neutrophils representing the relation of an area under a curve of neutrophilic granulocyte chemoluminescence induced by Staphylococcus epidermidis bacteria to an area under a curve of spontaneous neutrophilic granulocyte chemoluminescence, and if the derived value is less than 1.47, a high level of antibacterial resistance is stated, while the derived values 1.47 and more shows a low level of antibacterial body resistance.
EFFECT: method is informative, precise, complies with the modern requirements for laboratory diagnostic techniques and allows predicting a nature, a clinical course period and bacterial complications in the patients of various groups.
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Invention refers to medicine, in particular to immunology, and can be used to assess nonspecific antibacterial resistance of the human body.
A known method for the determination of nonspecific antibacterial resistance [EN 2068997 C1, publ. 10.11.1996]. The essence of the invention lies in the fact that prior to the preparation of the smear preparing a culture of the bacteria Serratia marcescens to use it as the object of phagocytosis, opsonized bacterial suspension serum of healthy individuals, incubated opsonization and Papanicolaou culture bacteria lacokanostra patients, examine the total cytochemical activity of myeloperoxidase and cationic proteins before and after incubation, and then calculate the stimulation index of neutrophils. When the index value of 0.5 and below define the reduction of antibiotic resistance of the organism. The disadvantage of this method is the duration of its execution and the need for microscopic evaluation of the activity of enzymes, which reduces the accuracy and objectivity of the method.
The objective of the invention is to create a new informative method for determining the antibiotic resistance of the organism.
The task is achieved by using a chemiluminescent analysis examined the functional activity of neutrophilic granulocyte the peripheral blood of the patient, determine the index of the bacterial activation of neutrophilic granulocytes (IBA-index), which represents the ratio of the area under the curve chemiluminescence of neutrophils induced by a bacterial suspension of Staphylococcus epidermidis to the area under the curve of spontaneous chemiluminescence of neutrophils. When the value of this index is below 1,47 determine the high level of antibiotic resistance, and when the amount equal to 1.47 and above, the low level of antibiotic resistance of the organism.
Low antibacterial resistance is characterized by increased susceptibility of the organism to bacterial infections; increasing duration of the disease and the accession opportunistic bacterial infections, as well as the weighting of the acute period and the rapid transition of the disease into a chronic form. Figure 1,47 obtained empirically based on the mapping data, a chemiluminescent analysis of the results of the medical examination and the subsequent exacerbations of chronic bacterial infections.
Activated neutrophilic granulocytes are potent effectors and trigger mechanisms of cascading reactions, ensuring the development of inflammation. Anti-infective action of neutrophils is associated mainly with the gene is a situation of active forms of oxygen, and one of the methods allowing to estimate oxygendependent biocenose neutrophilic granulocytes, is a chemiluminescent analysis. Neutrophilic granulocytes bear on their surface a wide range of receptors, some of which can interact with Neopleistocene bacteria (CR - and Toll-receptors). Phagocytosis without opsonization is associated with the presence of microbes specific virulence factors with which they penetrate into the cells. Phagocytosis is one of the main systems protect the body from foreign agents and plays an important role in inflammatory diseases, especially in staphylococcal infections. The undoubted advantage of this method is used as the inductor bacteria Staphylococcus epidermidis, which can cause severe inflammatory diseases.
The method is as follows. From the venous blood of the patient allocate neutrophilic granulocytes. For this to 5 ml of blood with heparin was added 1 ml of poliglyukina. The mixture is incubated for 30 min at 37°C to accelerate the sedimentation of erythrocytes. Received leukocyte supernatant washed twice in Hanks solution without phenol red for 10 min at 400 g. The supernatant is drained, the remaining neutrophilic granulocytes diluted in 1 ml of Hanks solution and get suspended. Count the number of natref is selected granulocytes in the camera Goryaeva. To conduct chemiluminescent analysis using the following reagents: donor serum (blood group AB, RH factor negative), Hanks solution (without phenol red), luminal concentration of 100 µg/ml Prepared sample: 200 ál suspension of neutrophils, 20 ál of donor serum, 240 μl of Hanks solution, 50 μl of lyuminola and 40 µl of bacterial suspension of Staphylococcus epidermidis in a concentration of 106CFU/ml Chemiluminescent analysis is performed in two cells: spontaneous chemiluminescence is carried out without adding the inductor, the second cell type bacterial suspension of Staphylococcus epidermidis. Measuring the chemiluminescent response exercise using a chemiluminescent analyzer, for example "CL3604"within 90 minutes recording the results and management of the chemiluminescent analyzer is performed via computer. Get the curves of spontaneous and induced bacteria chemiluminescence. The index of the bacterial activation of neutrophils (IBA-index) is determined by the formula:
Sinduced- the value of the area under the curve chemiluminescence induced bacterial suspension of Staphylococcus epidermidis;
Sspontaneous- the value of the area under the curve of spontaneous chemiluminescence.
When IBA-index below 1,47 diagnostics which can be carried on a high level of antibiotic resistance, when IBA-index equal to 1.47 or higher, the low level of antibiotic resistance of the organism.
The method was tested on 12 patients with chronic sinusitis, received treatment in the ENT Department of the Noosa heads "Road clinical hospital" on secretness JSC Russian Railways, and 45 healthy individuals. The survey results presented in table 1.
The results of the study found that all of 45 healthy individuals value IBA-index below 1,47 (100% match). In 11 patients also noted the coincidence of the results of the analysis. In 1 patient index value below 1,47. Thus, the coincidence method for determination of antibiotic resistance in patients with chronic sinusitis 91.7%. In the statistical analysis (with count median (Me) and the interquartile spread (C25-C75)) statistically significant differences (P<0.001) in the level of IBA in the groups of healthy and sick people (table 2).
|The level of IBA in the groups of healthy and sick persons|
Clinical example 1. Patient I., 43, case History No. 2149. Received in-patient treatment in the ENT Department of the Noosa heads "Road clinical hospital at the station Krasnoyarsk OJSC Russian Railways with a diagnosis of chronic rhinosinusitis. At admission the patient's condition moderate, concerned about the weakness and headaches. A study by the claimed method. IBA equal to 1.96 (above 1,47). Therefore, we examined the low level of antibiotic resistance. Clinical example 2. Patient S., 41, was surveyed all major medical specialists, including physician specialist physician, immunologist, when carrying out routine inspection of employees of JSC "Gorelektrosvyaz" on the basis of clinical research Institute of medical problems of the North SB RAMS. Deemed healthy. A study by the claimed method.
IBA 1.15 (below 1,47). Therefore, in subjects with a high level of antibiotic resistance.
The technical result of the proposed method:
- automated determination of the functionality of the activity of neutrophilic granulocytes;
- evaluation of the results does not depend on subjective factors.
Thus, the way of informative, accurate, meets all modern requirements for laboratory diagnostic techniques, allows to considerably simplify the process of determining factors of nonspecific defense, its functional activity and reserve capacity, makes it possible to identify the failure of phagocytic protection and its reactive or pathological activation.
The method of determining antibiotic resistance of the organism when the diseases caused staphyloccocal infection, based on the study of blood neutrophils, characterized in that the functional activity of neutrophilic granulocytes in the peripheral blood of the patient are examined using chemiluminescent analysis, determine the index of the bacterial activation of neutrophils (IBA-index), which represents the ratio of the area under the curve induced by the bacteria Staphylococcus epidermidis chemiluminescence of neutrophils to the area under the curve of spontaneous chemiluminescence of neutrophils, and when the value of this index is below 1,47 determine the high level of antibiotic resistance, and when the value is equal to 1.47 and up - low level of antibiotic resistance of the organism.
SUBSTANCE: method for prediction of rate of acute infectious diseases in infants with determining the immune status values on the first and fifth postnatal day: absolute lymphocyte count, differentiation marker carriers CD3, CD4, CD8, CD 19, CD56, CD95, a spontaneous nitroblue tetrazolium test value (NBTsp), a level of a phagocytic coefficient (PC), blood serum immunoglobulin A, M, G (IgA, IgM, IgG) concentrations, and further calculating a forecasting index (Ifor) with using a certain equation of multiple linear regression if Ifor does not exceed 3.2, a probability of no more than one acute infectious disease in an infant is concluded; if Ifor is within 3.2-3.7, 2 to 5 probable acute infectious diseases in an infant is stated; if Ifor exceeds 3.7, probability of more than 5 acute infectious diseases in an infant is concluded.
EFFECT: higher prediction accuracy.
SUBSTANCE: complete blood count is performed, and the leukogram components are used to calculate integral hematological values: Calph-Caliph leucocytic intoxication index - LII1, Ostrovsky leucocytic intoxication index - LII2, stress index - SI, neutrophil-lymphocyte index - NLI, neutrophil-monocyte index - NMI, lymphocyte-monocyte index - LMI. If observing the values LII1≤2.5, LII2≤2.75, SI≤1.0, NLI≤11.0, NMI≤16.0, LMI≤2.0, a mild severity is detected. The values LII1 2.6 to 4.5, LII2 2.76 to 5.0, SI 1.1 to 2.0, NLI 11.1 to 15.0, NMI 16.1 to 24.0, LMI 2.1 to 2.5 enables to predict a moderate severity level. A severe level is ensured by LII1> 4.5, LII2> 5.0, SI> 2.0, NLI> 15.0, NMI> 24.0, LMI> 2.5.
EFFECT: use of the declared method allows higher objectivity of determining severity level of acute pancreatitis.
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SUBSTANCE: method of early solid malignant disease staging (at the stage of provisional clinical diagnosis) by cell number with stable ontogenetic disorders detected in patient's peripheral blood lymphocytes by quantitative and qualitative analysis of stable chromosomal and genomic disorders in metaphase plates from patient's peripheral blood lymphocytes.
EFFECT: method enables following disease change shift in response to treatment and determining clinical effectiveness.
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SUBSTANCE: method of diagnosing rejection of kidney allotransplant includes determination of phase height PH of peripheral blood live lymphocytes by method of phase-interference microscopy, determination of quantity of lymphocytes with phase height PH≤1.5 mcm, 1.5 mcm<PH≤2 mcm, 2 mcm<PH≤2.5 mcm, PH>2.5 mcm, selection of lymphocyte activity coefficients for each limit, said phase heights of lymphocytes equal k3=3, k2=2, k1=1, k0=0 respectively. Obtained data are used to determine functional activity of lymphocytes in sample by formula: FA=(k3n3+k2n2+k1n1+k0n0)/n, where n is number of lymphocytes in sample, n3 is number of lymphocytes with PH≤1.5 mcm, n2 is number of lymphocytes with 1.5 mcm<PH≤2 mcm, n1 is number of lymphocytes with 2 mcm<PH≤2.5 mcm, n0 is number of lymphocytes with PH>2.5 mcm, k3, k2, k1, k0 are coefficients of lymphocyte activity, and if value of lymphocyte functional activity is within FA=1.8-2.0, rejection of kidney transplant is diagnosed.
EFFECT: application of claimed method makes it possible to set diagnosis in due time, which considerably increases efficiency of anti-crisis therapy in post-transplantation period.
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SUBSTANCE: claimed is method of diagnosing antioxidant function of membranes of erythrocytes in case of exacerbation of herpes-viral infection. Method lies in determination of superoxide dismutase concentration and titre of antibodies to herpes virus by method of immunosorbent assay (ELISA) of peripheral blood in pregnant women after exacerbation of herpes-virus infection. If superoxide dismutase concentration is 297.63±13.16 Unit/gHb (control - 386.77±13.12 Unit/gHb), titre of antibodies to herpes virus in peripheral blood of pregnant woman constituted 1:3200. If titre of antibodies is 1:6400, superoxide dismutase concentration decreased to 243.85±7.71 Unit/gHb (control - 386.77±13.12 Unit/gHb; p<0.001).
EFFECT: reduction of superoxide dismutase concentration in erythrocytes of peripheral blood in pregnant women during exacerbation of herpes-virus infection predicts reduction of protective reaction of toxic impact of lipid peroxidation on membranes of erythrocytes, increasing risk of their destruction.
SUBSTANCE: essence of method of predicting character of progressing course of chronic kidney disease (CKD) lies in determination of blood coagulation characteristics: activated partial thromboplastin time (aPTT), prothrombin time (PT), thrombin time (TT), antithrombin III (ATIII) level, von Willebrand factor (WF), endothelin-1 (E-1), D-dimer. If aPTT indices are within the range from 28 to 35 seconds, PT is from 12 to 15 seconds, TT is from 9 to 13 seconds, ATIII level is from 29 to 70%, WF level is from 105 to 115%, E-1 does nor exceed 0.32 fmol/ml and D-dimer is from 245 to 520 ng/ml FEU slow progressing of CKD is predicted. If aPTT, PT, TT, ATIII level are the same, and WF is higher than 115%, E-1 is higher than 0.32 fmol/ml, D-dimer is higher than 620 ng/ml FEU, accelerated CKD progressing is predicted.
EFFECT: increase of accuracy and reliability of chronic kidney disease diagnostics.
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SUBSTANCE: method includes determination of ceruloplasmin level on blood plasma. If its values are from 202.8 to 266.8 mg/l, thymus weight is higher than 0.2 g and content of lymphoblasts in it is more than 19%, development of re-inoculated lymphoblastic leukemia in line AKR/JY mice is diagnosed.
EFFECT: method makes it possible to diagnose syngenic re-inoculated lymphoblastic leukemia in mice quickly and accurately.
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SUBSTANCE: coagulation profile parameters are evaluated: fibrinolytic activity (FA), activated partial thromboplastin time (APTT), fibrin split products (D-Dimers) and international normalised ratio (INR). It is followed by calculating an integral coagulation factor (ICF) by formula: ICF=0.06FA+0.25INR+0.08 D-Dimer+0.54 APTT+6.54 where ICF is integral coagulation factor, FA is fibrinolytic activity (min.), INR is international normalised ratio (unit), D-Dimer is fibrin split products (ng/ml), APTT is activated partial thromboplastin time (sec.). The value ICF≥1 refers to a high risk, while the value ICF<1 - to a low risk.
EFFECT: application of the method enables evaluating a degree of risk of gestational thrombogenic complications that makes it possible to plan and control therapeutic actions aimed to normalise blood circulation in the mother-placenta-foetus system.
SUBSTANCE: in a cell containing an agitated blood sample, a primary measuring channel is formed by means of a collimated narrow optical beam. Then a photoconverter is used to register light intensity through the primary and to isolate thereafter constant and variable electric signal components, and to determine a mean value of thrombocyte aggregates. Immediately before completion of sample agitation, a mean ICK light intensity through termination the primary measuring channel formed in the cell at the bottom of the sample. On completion of agitation, an additional measuring channel similar to the primary one is formed at the top of the sample near to its level wherein time-stable light intensity value Iy is registered. A logarithm of the relation Iy/ICK shows a degree of aggregation.
EFFECT: invention enables measurement of the same sample both of mean aggregate value, and the degree of thrombocyte aggregation, and more accurate evaluation of these characteristics.
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SUBSTANCE: pressure equal to existing capillary pressure required to pass the entire red cell suspension by micro-sieving through a calibrated filter is generated in a simulated medium. It involves filtration time recording of the analysed blood by a stop watch. Passage time of the analysed red cell suspension shows a red cell deformability index - RCDI which is calculated by formulae: and where Xi is an average filtration time, n is number of observations.
EFFECT: use of the method allows quick and precise quantitative analysis of red cell deformability.
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SUBSTANCE: invention relates to laboratory methods for blood analysis. Plasma is dropped in copper sulfate solution with density 1.023 g/cm3, not above, and time for drop falling on bottom of graduated cylinder with column height 243 mm is measured. The blood plasma density value is calculated by the formula:
wherein is the unknown blood plasma density (g/cm3); is copper sulfate solution density measured by areometer (g/cm3); t is average falling time of plasma drop in the copper sulfate solution (as seconds); 0.260130126 and 0.00290695 are correction coefficients. Temperature of plasma and copper sulfate solution is 20oC. Method is simple and suitable and allows carrying out analysis of small volumes of blood plasma and to reduce analysis time.
EFFECT: improved assay method.
SUBSTANCE: method involves carrying out microscopic examination of blood serum samples taken from femoral vein and cubital vein. Femoral vein sample is taken on injured side. The examination is carried out before and after treatment. The blood serum samples are placed on fat-free glass slide in the amount of 0.01-0.02 ml as drops, dried at 18-30°C for 18-24 h. The set of pathological symptoms becoming larger or not changed after the treatment in comparison to sample taken before treatment, and morphological picture of samples under comparison taken from the cubital vein showing no changes or being changed to worse, the treatment is considered to be effective.
EFFECT: enabled medicamentous treatment evaluation in course of treatment to allow the treatment mode to be changed in due time; avoided surgical intervention (amputation); retained active life-style of aged patients.
FIELD: medicine, obstetrics, gynecology.
SUBSTANCE: in the first trimester of pregnancy one should study the content of CD8+CD11b lymphocytes and at their values being either equal or above 2% it is possible to predict gestosis. The present innovation enables to choose correct tactics of treating pregnant women that, in its turn, leads to decreased frequency of this complication of pregnancy and the risk for the development of fetal and neonatal pathology.
EFFECT: higher accuracy of prediction.
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SUBSTANCE: the present innovation deals with studying and treating diseases of inflammatory, autoimmune and degenerative genesis. One should perform sampling of heparinized blood followed by its sedimentation to obtain blood plasma with leukocytes and centrifuging to isolate the latter which are washed against erythrocytic and serumal admixtures, and, also, it deals with calculating the number of cells in samples out of leukocytic suspension after incubation (B) for 1.5 h at 37 C in holes of plastic microplotting board, out of leukocytic suspension one should additionally prepare two samples, one should be applied to calculate total number of leukocytes before incubation (A), the second sample undergoes incubation at the same mode at addition of autoserum to calculate the number of cells remained after incubation (C). One should state upon adhesive properties of leukocytes by the index of spontaneous adhesion (D), where D=(A-B)/B.100%, and effect for enhanced cellular adhesion under the impact of autoserum should be detected by the value of K=(B-C)/C.100% at K ≥ 30%, where B - C - the number of cells undergone additional adhesion after addition of autoserum. The present innovation widens functional possibilities of the suggested method due to obtaining additional values depicting adhesive properties of blood leukocytes.
EFFECT: higher accuracy of detection.
FIELD: medicine, diagnostics.
SUBSTANCE: the present innovation deals with blood sampling, separating plasma against erythrocytes, moreover, in plasma on should detect activity of antithrombin III, proteins C and S, XIIa-dependent fibrinolysis and concentration of plasminogen obtained results should be expressed as relative units followed by calculating integral parameter that characterizes the state of anticoagulant-fibrinolytic potential (IPAFP) by the following formula: IPAFP = [(C1 + C2)/(C3 + C4)] x 100, where C1 - the ratio of observed value of antithrombin III activity to the value of inferior border of the range of analogous parameter norm; C2 - the ratio of observed value for the activity of proteins C and S system to the value of inferior border of the range of this parameter norm; C3 - the ratio of the value of inferior border of plasminogen concentration under normal conditions to observed value of analyzed parameter; C4 - coefficient calculated with the help of regression equation: C4 = 0.9 + (0.01 x X), where X - terms of lysis of patient's euglobulin clot/min, and at IPAFP value of 101.4 U and higher one should state anticoagulant-fibrinolytic blood potential to be in norm, in interval of 64.8 - 101.3 -as insufficient, and at 64.7 and below - as critical. The present method simplifies the procedure of evaluating the state of endogenous anticoagulants and activity of XIIa-dependent fibrinolysis.
EFFECT: increased diagnostic value of obtained results.
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FIELD: medicine, laboratory diagnosis.
SUBSTANCE: method involves determination of the patient blood content of globulin-alpha 1, globulin-beta, globulin-gamma and the total bilirubin content followed by calculation of diagnosis indices for the patient (Y1, Y2, Y3) by using the computer program "Statistica 1.5" and introducing values X1, X2, X3 and X4 in computer wherein X1 means globulin-alpha 1 value; X2 means globulin-beta value; X3 means globulin-gamma value; X4 means total bilirubin value. Obtained values of diagnosis indices for the individual patient (Y1, Y2, Y3) are compared with average values of diagnosis indices (Y1', Y2', Y3') for different urogenital infections followed by comparison by sign and value. By the maximal coincidence of diagnosis index values for the individual patient with average diagnosis index values urogenital disease is diagnosed and the following diagnosis index average values are used: for chlamydiosis: Y1' = -2; Y2' = -0.1; Y3' = -0.2; for mycoplasmosis: Y1' = 2; Y2' = 0.8; Y3' = -0.04; for ureaplasmosis: Y1' = 2; Y2' = -1; Y3' = 0.02; for health persons: Y1' = -2; Y2' = 0.1; Y3' = 0.2. Invention provides the development of a method for express-diagnosis of infection at initial stage and diagnosis of atypical forms that occur in these diseases, and differential diagnosis of chlamydiosis, mycoplasmosis and ureaplasmosis. Invention can be used for carrying out the differential diagnosis of chlamydiosis, mycoplasmosis and ureaplasmosis.
EFFECT: improved method for express-diagnosis.
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SUBSTANCE: method involves determining absolute value of ratio between lymphocyte number and absolute value of monocyte number in peripheral blood at the end of combine radiation therapy. The ratio is divided by 4.05. The result value being greater than 1, no disease relapse occurrence is predicted during the first observation year. The value being less than 1, tumor growth progress is stated and carcinoma relapse is predicted at the first year after treatment.
EFFECT: enhanced accuracy in detecting pathological process progress before observing clinical manifestations.
SUBSTANCE: method involves determining infrared radiation absorption coefficient in blood plasma in bandwidth of 1543-1396 cm-1. The infrared radiation absorption coefficient is determined in %. The value being equal to 29.7±1.1%, catarrhal cholecystitis is diagnosed. The value being 26.4±1.4%, phlegmonous cholecystitis is diagnosed. The value being 21.2±1.8%, gangrenous cholecystitis is diagnosed. The value being equal to 18.6±0.5%, gangrenous perforated cholecystitis case is diagnosed. The value in norm is equal to 32.4±0.8%.
EFFECT: high accuracy and specificity of diagnosis.
SUBSTANCE: the present innovation deals with biomedical measuring technologies, in particular, to those to detect bactericide activity of blood serum according to the level of its inhibiting impact upon luminescence intensity of sulfur-sensitive luminescent bacteria (ΣimpO) against control - luminescence intensity the same sulfur-sensitive luminescent bacteria that had no contact with blood serum (ΣimpK), then one should calculate the value of bactericide activity of blood serum by the following formula:
As sulfur-sensitive luminescent bacteria one should apply either natural or recombinant microorganisms being characterized by direct proportionality between intensity of decreased spontaneous bioluminescence level and degree of bactericide effect. For example, it is possible to apply Escherichia coli strain with genes of Photobacterium leiognathi luminescent system. The suggested method enables to shorten the duration for detecting bactericide activity of blood serum and decrease its labor intensity.
EFFECT: higher efficiency of detection.
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SUBSTANCE: method involves determining blood insulin I and thyroxin T content and phagocytic leukocyte activity (PLA). Activity coefficient is calculated on the basis of formula KA=IxPLA/T. KA value being found greater than 2.8 units, considerable amelioration treatment effect is predicted. The value being from 1.4 to 2.8 units, amelioration is predicted. KA being less than 1.4 units, lower treatment efficiency is predicted.
EFFECT: high reliability of prognosis.