Method for diagnosis of ehrlich's carcinoma
FIELD: medicine and biochemistry.
SUBSTANCE: claimed method includes blood lymphocyte treatment with labeled fluorescein isothiocyanate of pea lectin isoform obtained by extraction of milled pea seeds with sodium chloride solution, purifying by undercooling, dialysis and isoform isolating by column chromatography on Sephadex, and elution with glucose-containing solution, and when number of labeled lymphocyte is lower than 50 % in contrast to normal one Ehrlich's carcinoma is diagnosed.
EFFECT: diagnosis method of increased specificity.
1 tbl, 3 dwg
The invention relates to medicine, in particular to methods of diagnosis of cancer.
One of the main problems of Oncology is that no one can find the flaws in the immune system of cancer patients, which make possible the growth of tumors in the body. There is a paradoxical situation when the immune system of cancer patients does not differ from healthy, but in the body are growing tumor.
On the background of tumor growth in immune system doesn't change, such as those that develop in immunodeficiency States, when depressed, entire groups of immunocompetent cells or when there is disappearance of any cell receptors. The changes that occur in lymphocytes of the body on the background of tumor growth, affect glycosylation of proteins (receptors) in their cell membranes. In this case, the immunocompetent cells lose their ability to respond only to the tumor cells, whereas the response to infectious agents fully preserved, i.e. does not develop immunodeficiency.
Currently, all methods of diagnosis of cancer blood test is based on measuring the level of tumor associated antigens. Opukholeobrazovanie antigens are patterns, which are normally presented in the tissues of the body the ISM, but their number in normal tissues or slightly, or the location of their tissues is that normal in the blood, they do not fall. As a result of development of oncological process increases the number of tumor cells and increases the body the number of tumor associated antigens that are expressed on the surface of these cells. When death and / or peeling from the surface of these tumor antigens are starting to get into the blood. Thus, this group of methods of diagnostics uses patterns that are on tumor tissues and completely ignores the changes that can occur in the body in response to the growing tumor.
There is a method of diagnosing cancer by determining the content of small lymphocytes (diameter <7.5 μm) in the blood of cancer patients (Viganello and other "Decrease in the content of small lymphocytes in the blood of patients with malignant bone tumours. Oncology issues. 1987, volume 33, No. 9, p.15-21).
As the authors state, reducing them to 40-75% compared with healthy people is observed on the background of growth of tumors.
However, this method has a great subjective and depends on the qualification of specialists. Thus, the diameter of the cells may vary depending on the isotonicity of the solution, the rate of fixation of smears, optizone sizes of cells eyepiece-micrometer MOU-1, etc. Due to the high subjectivity of the method he has not found wide application.
The objective of the invention is to increase the specificity of diagnosis of cancer and preventing its subjectivity.
The problem is solved by the method lies in the fact that the blood lymphocytes treated with labeled fluorescent dye - fluoresceinisothiocyanate isoform of pea lectin, and by reducing the number of labeled lymphocytes by more than 50% with respect to the norm judge the presence of cancer.
The method is as follows.
Lectin peas allocate the method of affinity chromatography on Sephadex G-100. Selection isoforms of pea lectin with high affinity binding to sugars is a necessary step to improve the specificity of diagnosis, because otherwise with the surface of the cells contacted with the lectin isoforms that when laundering cells from unbound lectin was also removed (due to their low binding constants) with the cell surface and this would lead to a reduction in the percentage of stained cells.
For this purpose, seeds of peas grind in the mill into flour with grain sizes less than 0.1 mm, Then pour 4 times by weight the amount of 0.5% of NaCl and infused for 3 hours at 4°C. the Extract is acidified to a pH of 4.6 with 5N HCl and centrifuged DL the removal of sediment at 1500 g for 30 minutes
Solid ammonium sulfate (NH4)2SO4added to the supernatant to achieve a 60% saturated solution. Precipitated precipitated protein is collected by centrifugation at 3000 g for 30 minutes the Precipitate is dissolved in a minimal volume of water and deleteroute strictly at 4°against M of NaCl in M phosphate buffer (pH 7.2). Protein precipitated precipitated during dialysis, is also removed by centrifugation at 3000 g for 30 minutes
The supernatant is applied on a column of Sephadex G-100 (26×100 cm). Unbound protein elute 4 l M of NaCl in M phosphate buffer (pH 7.2). Lectin peas elute M of NaCl in M phosphate buffer (pH 7.2) containing glucose from 0.01 to M, with a step increase in glucose concentrations 0.01, 0.05 and M. Suirvey lectin collected in three separate fraction, which is subjected to dialysis against M of NaCl in M phosphate buffer (pH 7.2). Protein is sterilized by filtration using 0.22μ Millipore filter.
Next, perform the tagging isoforms lectin fluoresceinisothiocyanate. Tagging isoforms lectin this fluorescent dye is necessary because it is fluoresceinisothiocyanate fluorescent from the laser light source, which is used in a flow cytometer.
This device combines together two block - optical device with a laser light source and power the second computer. Cells of the analyzed sample are moving in a thin glass capillary, a laser beam shines through the capillary and the cell that it can move. In different directions along the laser beam are numerous detectors, which measure the passage of light through the cell, its dispersion on the object, reflection, fluorescence, etc.
A computer with a special program evaluates the received information and displays the optical parameters of each, held by capillary cells, determining the level of fluorescence of each of them that allows you to measure the amount of lectin, contacting the cell and, consequently, the level of sugars on the surface of the analyzed cells that are available for binding with the lectins.
The original solution of lectin stored in M NaCl in M phosphate buffer (pH 7.2) with addition of sodium azide to a final concentration of 0.1% at a concentration of 8 mg of protein per ml For labeling take 0.5 ml of the solution and bring the same buffer volume to 2.5 ml of This volume applied to the column PD-10, balanced 0.1M carbonate buffer pH 9.3. Protein elute 3 ml of carbonate buffer. Thus obtained protein solution with a concentration in the range of 0.5-2 mg/ml 1 mg fluoresceinisothiocyanate dissolved in 1 ml of dimethyl sulfoxide and added to the protein solution at the rate of 25 µl per 1 ml of buffer. The solution peremeci is try and leave overnight at 4° C. after incubation, the solution was added 100 μl of 0.5M solution of ammonium chloride and Stavelot in the same terms if 4°With 2 hours. Then lectin translate in the source buffer (M of NaCl in M phosphate buffer (pH 7.2) with addition of sodium azide to a final concentration of 0.1%) using a column PD-10, balanced M of NaCl in M phosphate buffer (pH 7.2) with addition of sodium azide to a final concentration of 0.1%.
Diagnosis of cancer at the transplantable Ehrlich carcinoma
Transplantable tumor models allow us to quickly explore the changes that occur in the body on the background of tumor growth. To do this, take the animals one line, weight, sex, age, and part of them perejivaut the tumor and part is used as a control. Most of the tumors after two to three weeks to reach considerable size, while animals without tumors are adequate control on the changes in the body that occur on tumor growth.
On the growth of Ehrlich carcinoma in the serum of mice appear for this tumor factors that accelerate its growth in in vivo studies. However, the nature of these changes remains unknown, as it is not possible to detect any tumor-specific factors. We have made the assumption that this phenomenon is associated with changes in the glycosylation of elkow blood and membranes of lymphocytes. To confirm this assumption was studied by changing the binding of the labeled lectin isoforms peas with blood leukocytes on the growth of Ehrlich carcinoma. Selected isoforms lectin has the ability to contact the glucoside and mannose, including part of the polysaccharides in the glycosylation of proteins.
The method is carried out using male mice F1(CBAxC57BL/6), mice of C57BL/6 males. Each group used 10 animals. Cells of Ehrlich carcinoma transplanted intramuscularly in a dose of 106cells in 0.2 ml of RPMI-1640 medium in the mouse. Lymphocytes isolated from the peripheral blood of mice on day 15 after transplantation of the tumor by a standard method in the density gradient 1.093. Each experiment is repeated at least 3 times.
Determination of the expression of surface markers on the surface of lymphocytes carried out using labeled fluoresceinisothiocyanate lectins. Evaluation of the binding specificity of pectins from the cell surface to confirm the inhibition of their binding adding sugars. The results take into account the method of running cytofluorometry on a flow cytometer FACScan (Becton Dickinson, USA). Gate (window) cell population set based on the combination of forward and side light scattering and size of cells. This approach completely eliminates subjecti the significant factor in the analysis, so the measurements are performed by the detectors and their readings are entirely determined by the optical properties of the analyzed object. When taking into account the results of counting 10,000 events in gate. Statistical processing of the material is done using a software package WINMDI 2.8.
Investigated the binding of lectins to the surface of peripheral blood lymphocytes intact mice and mice on the 20th day of Ehrlich carcinoma. Table 1 shows the averaged data for all series of experiments with different visible tumor volume from a minimum of less than 1 mm3up to 5 cm3.
The figure 1 shows the distribution of cells in the blood lymphocytes of healthy mice. The abscissa axis shows the distribution of cells of peripheral blood lymphocytes depending on their front light scattering, and the ordinate axis shows the light scattering of cells depending on their lateral light scattering. Lymphocytes form a dense cloud, which indicates the homogeneity of the considered cells. The selected frame (polygon) in figure 1 is called the gate, it shows that the population of lymphocytes in the blood of mice, which are studied for linking them with lectins. The analyzed cells have similar optical characteristics as to control animals and animals-carriers of tumour, which also excludes the subject is wny factor in the research.
The figure 2 presents a histogram of the distribution of normal blood leukocytes before painting pea lectin (histogram 1) and after painting FITZ-labeled lectin peas (histogram 2). Therefore, after incubation with the lectin shift the histogram to the right along the x-axis, indicating that the increase in fluorescence of the cells due to binding with them labeled lectin isoforms. The percentage of connected cells is to 91.2%. The ordinate axis shows the number of cells that have the appropriate level of fluorescence.
Figure 3 shows histograms of lymphocytes in the peripheral blood of animals-carriers of tumour to their color isoform of pea lectin and after painting. Just as in the case of normal lymphocytes, the color of the pea lectin causes a shift of the histogram to the right. Only 21.3% of cells are in the same color, whereas in the control of such cells to 91.2%. Thus, the percentage of labeled cells decreased by 69,9% (91,2-21,3=69,9).
Thus, the color labeled lectin isoform pea peripheral blood lymphocytes of mice showed that despite the growth of tumors, a decrease in the availability of sugar (glycoside and mannose) for binding to lectins on the cell surface. The changes of glycosylation of membrane proteins can cause damage to the invasive properties of the cells and the reduction of binding them to the surface of tumor cells, that can lead to failure of antitumor immunity.
Allocated in accordance with the invention isoform of pea lectin can be used for the diagnosis of cancer (see table).
|The average percentage of labeled cells in animals without tumors||The average percentage of labeled cells in animals with tumors||The reduction in the percentage of labeled cells on the background of tumor growth|
|(in %)||(in %)||(in %)|
A method for diagnosing carcinoma, Ehrlich by studies of blood lymphocytes, characterized in that the blood lymphocytes treated with labeled fluoresceinisothiocyanate isoform of pea lectin obtained by extraction of the crushed seeds of peas solution of sodium chloride, purification presidenial, dialysis and allocation isoforms column chromatography on Sephadex and elution with a solution containing glucose, and by reducing the number of labeled lymphocytes by more than 50% with respect to the norm diagnosed with Ehrlich carcinoma.
SUBSTANCE: invention relates to method for biological material sample analysis in biological or immunological tests, wherein in sample treatment process coloring is observed, and color darkening is correlated with amount of tested substance. At least one color characteristic, such as coloring angle, chromaticity, darkening, and obtained color brightness is measured. Method of present invention is useful in analysis of lung, throat, cervix uteri or spermatic liquid mucilage for diagnosis of cancerous and precancerous conditions.
EFFECT: enhanced assortment of agents for diagnosis of cancerous and precancerous conditions.
14 cl, 4 dwg
SUBSTANCE: method involves determining lymphocyte enzyme activity. Blood sample is taken and NAD-dependent glutamate dehydrogenase, malate dehydrogenase activity is measured. Combination of both dehydrogenases activity is interpreted in terms of cardiac rhythm disorders. The combination of glutamate dehydrogenase activity from 3.37 to 5.62 mcU and malate dehydrogenases activity from 9.55 to 17.60 mcU points out to combined cardiac rhythm disorder. The combination of NAD-dependent glutamate dehydrogenase activity from 5.63 to 10.37 mcU and malate dehydrogenases activity from 17.61 to 26.42 mcU points out to cardiac rhythm disorder caused by pulse formation disorder.
EFFECT: high diagnosis objectivity level.
FIELD: medicine, surgery, biochemistry.
SUBSTANCE: in erythrocytes of peripheral blood in patients with colorectal cancer one should detect the activity of total ATP-ase and Na+-K+-ATP-ase on the 4th d after operation. At increased activity of total ATP-ase to the 4th d being below 0.6 mcM/h per 1 mg protein and Na+-K+-ATP-ase being below 0.3 mcM/h per 1 mg protein it is necessary to predict the development of acute gastroduodenal ulcers. The innovation leads to more simplified technique applied.
EFFECT: increased accuracy of prediction.
SUBSTANCE: method involves determining functional monocyte activity in spontaneous and stimulated nitroblue tetrazolium (NBT) test. Diagnostic index Kst is calculated as spontaneous-to-stimulated NBT-test values ratio. Kst being equal to or greater than 1.5, newborn adaptation process is considered to be adequate, otherwise, early adaptation period disorders and functional exhaustion are to be diagnosed at the level of the immune system monocyte chain.
EFFECT: high accuracy in revealing early stage pre-clinical signs.
FIELD: medicine, oncology.
SUBSTANCE: in plasma of peripheral blood in children one should detect cathepsin D activity and antiproteolytic activity and calculate coefficient of their ratio. At obtaining the value of coefficient being equal 10.5 ± 2 one should detect lymphadenopathy of benign genesis in a child to choose a waiting-type tactics at prescribing antiphlogistic therapy. At coefficient value ranged 30-100 on should detect malignant lymphoproliferative process in a child to immediately fulfill an excision biopsy of a lymph node along histological studying and at proving the diagnosis special therapy should be indicated. Application of the present method enables to conduct additional objective differential-diagnostic test under disputable cases at the stage being before carrying out histological analysis of lymph nodes and prescribe necessary therapy for every concrete case in due time.
EFFECT: higher accuracy of differential diagnostics.
3 ex, 1 tbl
FIELD: biochemistry, biotechnology.
SUBSTANCE: claimed method includes sample treatment with alkali copper-containing reagent comprising 49 pts of 2 % sodium carbonate solution in 2 N sodium hydroxide (A) and 1 pts of 0.5 % blue copper in 3.33 % sodium or potassium tartrate followed by addition of Folin's reagent and mixture conditioning in ultra-thermostat at 50°C for 10 min. Method of present invention allows reducing analysis time to 20 min and increasing sensitivity and reproducibility protein content determination by Lowry method.
EFFECT: accelerated method for determination of protein content in biological liquids and enzyme solutions.
2 dwg, 1 tbl, 3 ex
FIELD: experimental medicine.
SUBSTANCE: on the 3d - 4th d of experimental staphylococcal acute otitis media it is necessary to examine lactoferrin concentration in nasal mucosa and activity of beta-lysines in blood serum in experimental rabbits and at lactoferrin concentration being 119.8 ± 1.6 ng/ml (standard being 86.2 ± 1.4 ng/ml) and activity of beta-lysines being 69.4 ± 0.4% (standard being 54.8 ± 1.7%) it is possible to conclude on acute phase of inflammatory process in patient's middle ear.
EFFECT: higher accuracy of detection.
SUBSTANCE: invention relates to unified assay of total flavonoids in three kinds of eyebright grass: Euphrasia brevipila Burnat@Gremli, Euphrasia parviflora Schag, and Euphrasia X reuteri Wettst, as well as in extractive preparations of Euphrasia brevipila Burnat@Gremli. Claimed method includes utilization of 2 % aluminum chloride alcohol solution as chelating agent and 8 % sodium acetate alcohol solution as ionizing agent; detection of colored complex by differential spectrophotometry at λ = 382±2 nm; and calculation of total flavonoids using specific absorption coefficient for cinarozide as reference standard and individual dilution series for each extractive preparation.
EFFECT: method for standardization of drug raw materials and eyebright extractive preparations according to active ingredient (flavonoid) content, useful in development of normative and technical documentation.
6 cl, 8 tbl, 6 ex, 1 dwg
FIELD: bioassay methods.
SUBSTANCE: invention, in particular, relates to a reagent system for detecting substances. Signal generation system for detecting substances in a sample contains at least one first and second electron transfer agent and redox indicator. Electron transfer agents utilized are those of protein and nonprotein nature, for example phenazine compound and diaphorase. System further contains at least one of following compounds: enzyme cofactor, enzyme manifesting oxidative activity relative to tested substance, e.g. corresponding dehydrogenase. Proposed systems, reagent compositions, test strips, and kits find use in detection of a large number of substances in a sample such as a biological sample, e.g. blood or blood fraction.
EFFECT: increased reaction rate of signal generation system and reduced cost.
8 cl, 1 dwg, 3 tbl
FIELD: medicine, neurology.
SUBSTANCE: in patient's blood serum on should detect concentrations of lactic and pyroracemic acids, activity of lactate dehydrogenase enzyme to calculate the coefficient C by the following formula C=(LA x LDG)/PRA x K, where K - a balance constant of corresponding dehydrogenase reaction, for cerebral tissue it corresponds to 99.18; LA - concentration of lactic acid, LDG - activity of lactate dehydrogenase enzyme; PRA - concentration of pyroracemic acid. At C value being up to 597.17 one should predict favorable flow of cerebral ischemia, at C values being above 597.17 - unfavorable flow. The innovation enables to objectively evaluate the state of redox processes in patient's body and based upon this information predict the flow of the disease mentioned that favors the prescription of adequate therapy.
EFFECT: higher accuracy and efficiency of prediction.
2 ex, 1 tbl
SUBSTANCE: method involves determining plasminogen, fibrinogen and fibrin degradation products level and blood platelets aggregation and disaggregation values in treating a patient. Blood platelets aggregation rate growing twice as high at the fifth day, blood platelets disaggregation growing 5-8 times as high at the third-fifth day, plasminogen concentration growing 1.5 times as high at the eighth day and fibrinogen and fibrin degradation products level dropping 2.5 times as low at the eighth day, favorable treatment outcome is to be predicted. Aggregation rate dropping twice as low at the fifth day with blood platelets disaggregation being lost at the eighth day and no fibrinogen and fibrin degradation products level changes being detected, unfavorable prognosis is considered to be the case.
EFFECT: higher accuracy of early stage prognosis.
SUBSTANCE: method involves determining gas parameters of blood in noninvasive way in rest state and under exercise stress without heart cavities and large blood vessels catheterization being applied. The obtained data are processed using calculation formulas.
EFFECT: higher accuracy in determining venous HbO2 mean value and arteriovenous oxygen difference in systemic circulation.
FIELD: clinical medicine, laboratory diagnostics.
SUBSTANCE: one should carry out complex laboratory survey including daily urinary sampling by Zimnitsky and blood sampling for analysis just after finishing sampling the last urinary portion. In every urinary portion one should detect pH, protein and relative density, in total urinary volume it is necessary to detect concentration and quantity of uric acid, creatinine and daily proteinuria, calculate clearances of uric acid and creatinine and fractional clearance of uric acid. On detecting purine exchange disorders including high concentration of urinary uric acid at low daily diuresis one should recommend to widen drinking mode; at altering urinary reaction towards acidic side one should prescribe alkalizing preparations; in case of pronounced purine exchange disorders and severe accompanied pathology one should carry out extracorporal therapy. The innovation enables to considerably improve the quality of diagnostics and match the most optimal schemes of therapy.
EFFECT: higher efficiency of therapy.
5 cl, 3 dwg, 5 ex, 20 tbl
FIELD: medicine, laboratory investigations.
SUBSTANCE: method for preparing a blood preparation for electron-microscopic investigation involves centrifugation of suspension of non-fixed isolated cells at the rate 1500 rev/min, removal of supernatant, resuspending a precipitate in 0.25 ml of 10% human serum albumin followed by addition one drop of 25% glutaric dialdehyde to the prepared mixture and centrifugation up to preparing gel. Prepared gel is poured with 10% formalin in phosphate buffer, pH 7.5, for 24 h followed by separation of gel for segments. Gel segments are subjected for additional fixation in 2% solution of osmium tetraoxide, dehydrated, poured in epoxy resins, slices are prepared, stained and contrasted. Method allows significant improving quality of the preparation.
EFFECT: improved preparing method.
2 cl, 3 dwg
SUBSTANCE: method involves studying blood serum with wedge dehydration. Blood serum drop is first dried on object-plate and the prepared facia is examined with microscope. Anomalous reticular radial structures being observed in central and/or intermediate zones of the facia, moderate hyperparaproteinemia degree is diagnosed. Isolated lumps and/or lumps located on crack periphery in central zones of the facia being observed, highly marked hyperparaproteinemia degree is diagnosed.
EFFECT: simplified accelerated method.
3 dwg, 1 tbl
SUBSTANCE: method involves analyzing whole capillary blood on monocytosis and lymphocytosis in the cases the depression is clinically apparent according to Hamilton scale at 22.1±2.3 points determining infrared blood spectroscopy absorption indices in infrared spectral analyzer during 30 s in bandwidth ranges of 3085-2832 cm-1, 1543-1425 cm-1, and 1087-963 cm-1, and determining mean absorption indices. The mean absorption indices being found equal to 40.2±2.0%, 38.0±3.2%, 43.1±1.9%, in bandwidth ranges of 3085-2832 cm-1, 1543-1425 cm-1, and 1087-963 cm-1, in combination with normal monocytosis and lymphocytosis, respectively, depression in remote mild craniocerebral injury period is to be diagnosed. The mean absorption indices being found equal to 49.5±2.6%, 54.4±3.2%, 47.8±2.8%, in bandwidth ranges of 3085-2832 cm-1, 1543-1425 cm-1, and 1087-963 cm-1, in combination with monocytosis > 5% and lymphocytosis >25% in leukocytic formula of bulk blood analysis, respectively, therapeutically resistant depression in remote mild craniocerebral injury period is to be diagnosed.
EFFECT: high accuracy of diagnosis.
SUBSTANCE: method involves studying urine gas discharge visualization parameters at the first days of life. To do it, urine drops are placed into magnetic field of high intensity with bipolar pulses amplitude being equal to 16 kV pulse succession frequency of 1024 Hz and pulse series duration of 0.5 s. Time required for performing single study being equal to 10 min. The gas discharge visualization picture parameters like fractal discretization being equal to 2.07±0.05, fragmentation 13.93±1.81 and brightness 224.6±0.59, favorable newborn state outcome is to be predicted. The gas discharge visualization picture parameters like fractal discretization being equal to 1.873±0.11, fragmentation 5.909±2.64 and brightness 222.8±0.65, unfavorable newborn state outcome is to be predicted. Cases of gas discharge visualization picture parameters like fractal discretization being equal to 2.128±0.08, fragmentation 21.17±2.123 and brightness 226.8±0.8 are considered to be norm.
EFFECT: simplified noninvasive test.
3 dwg, 1 tbl
FIELD: medicine, in particular laboratory diagnosis of hemopexis system damage useful in anticoagulant therapy control.
SUBSTANCE: hematocrit is determined in capillary blood of subject; then PT value is twice measured, and mean value is calculated. Said mean value is correlated with PT value of normal plasma and INR is calculated according the formula including coefficient which depends on hematocrit value and International index of thromboplastin reagent sensitivity.
EFFECT: standardized PT value-based method for INR value determination of improved accuracy.
2 cl, 1 tbl, 3 ex, 1 dwg
SUBSTANCE: method involves scanning pulse with glass plate placed on pulse measurement point. The glass plate bears biological test system containing composition of 0.1% aqueous amino acid solutions like tryptophan, aspartic acid, alanine, treonine, valine, serine, glycine and tyrosine taken in equal proportions, 0.5% aqueous solution of dopamine, 0.5% aqueous solution of histamine, 12% aqueous solution of magnesium sulfate taken in 3:3:13 proportion. The plate is hold on pulse for 1-2 min before and 10-15 min after giving the pharma- or parapharmaceutic under study to a patient. Then, the plate is dried at T=+18-20°C during 2-3 min and another glass plate is covered with biological test system surrounded with wall of the pharma- or parapharmaceutic under study arranged along periphery and dried at T=+18-20°C during 2-3 min. The biological test system structures formed on the plates are compared in polarized light with quartz compensator. Structural similarity being detected, identification of pharma- or parapharmaceutic introduced into human organism is to be recorded.
EFFECT: enhanced effectiveness in identifying biologically active substances introduced into human organism.
FIELD: veterinary science.
SUBSTANCE: the present innovation deals with applying reaction of agglutination inhibiting including dilution of test-sera and tested sera with sodium chloride physiological solution followed by their incubation at 37° C, performing reaction of hemagglutination due to double serial dilutions, mixing tested serumal dilutions with a diagnostic kit. Then comes repeated incubation at 37° C to detect the index of inhibition, moreover, reaction of hemagglutination due to the method of double serial dilutions should be fulfilled on a plot and at the index of inhibition being either above or equal to 1.1 it is possible to diagnose secondary immunodeficiency. The innovation enables to decrease labor capacity of the method described, enables to detect all immunodeficient animals and adapt this method to mass trials of blood sera in the field of veterinary science.
EFFECT: increased sensitivity of the method.
1 dwg, 1 tbl
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.