Reagent for total α-amylase activity test
SUBSTANCE: reagent for testing total α-amylase activity in serum, blood plasma and urine contains as a substratum 2-chlorine-4-nitrophenl-4-O -β-D-galactopyranosylmaltoside (GalG2CNP), sodium chloride, potassium sulphocyanate, sodium azide, EDTA, calcium acetate, Triton X-100, 2-morpholinoethanesulfonic acid (MES) and water. The pH value of said reagent is 5.5-6.5.
EFFECT: use of the reagent enables high precision and results reproducibility α-amylase test.
1 tbl, 4 ex
The invention relates to the field of medicine and biochemistry, namely the creation of reagents, allowing to determine the activity of the total α-amylase in serum, plasma and urine.
For determining the activity of α-amylase various techniques have been developed, based on the ability of the enzyme to hydrolyze internal α-1,4-glycosidic bonds oligo - and polysaccharides.
The most common use for determining the activity of total α-amylase is a method comprising using as the substrate 4,6-ethylidene-(G7)-p-nitrophenil(G1)-α,D-maltoheptaoside (4,6-Tilden(G7)-p-nitrophenyl-(G1)-α,D-maltoheptaoside; Et-G7PNP)approved by the International Federation of clinical chemistry (IFCC). In the reaction of α-amylase cleaves Et-G7PNP to G2PNP, G3PNP, G4PNP, Et-G3Et-G4and Et-G5. Then α-glucosidase hydrolyzes G2PNP, G3PNP and G4PNP with the formation of glucose and colored substance p-NITROPHENOL. The rate of accumulation of colored product is directly proportional to the activity of total α-amylase in the sample (Kruse-Jarres JD, Kaiser S., Hafkenscheid J... et al. Evaluation of a new α-amylase assay using 4,6-ethylidene-(G7)-l-4-nitrophenil(G1)-α-D-maltoheptaoside as substrate // J. Clin. Chem. Clin. Biochem. 1988. Vol.27. P.103-113).
The main disadvantage of this method is the two-phase process of the reaction and the use of additional enzyme for p which produce a colored product.
Closest to the claimed reagent prototype is a set of reagents for determining the activity of total α-amylase kinetic colorimetric method, which consists of two reagents, the following compounds: reagent 1 - sodium chloride 50-500 mmol/l, potassium radamisty (KSCN) 50-400 mmol/l, sodium azide 1-90 mmol/l, calcium chloride 0.5 to 10.0 mmol/l MES (2-morpholinepropanesulfonic acid) 50 mmol/l, water - the rest, the pH of the reagent 5-8; reagent 2 containing: 2-chloro-4-nitrophenyl-4-O-β-D-galactopyranosylmaltoside (2-chloro-4-nitrophenyl-4-O-β-D-galactopyranosyl; GalG2CNP) 2,4-12 mmol/l MES, 50 mmol/l, water(rest)pH reagent 5-8. The basis set for determining the activity of total α-amylase laid kinetic colorimetric method, which consists in the fact that the α-amylase hydrolyzes GalG2CNP with the formation of a colored product 2-chloro-4-nitrophenol and unpainted - 4-O-β-D-galactopyranosylmaltose. The rate of accumulation of the colored product is directly proportional to the activity of α-amylase in the sample (Morishita Y., IinumaY., Nakashima N., et al. Total and pancreatic amylase measured with 2-chloro-4-nitrophenyl-4-O-β-D-galactopyranosylmaltoside // Clinical Chemistry. 2000. Vol 46. P.928-933).
The disadvantages of the prototype are:
- small shelf life of reagents: 3 months at 4°C;
- the complexity of using the system of the two reagents.
An object of the invention is to increase the stability of the reagent for measuring the population activity total α-amylase in serum, plasma and urine with high accuracy and reproducibility of the analysis results.
The goal of the project is achieved by the proposed reagent containing the following components: a substrate GalG2CNP 0.5 to 20 mmol/l, sodium chloride 50-500 mmol/l KSCN 50-400 mmol/l, sodium azide 1-90 mmol/l, EDTA (ethylenediaminetetraacetic acid) 0.2 to 5.0 mmol/l, Triton X-100 0,075%vol., calcium acetate 0.7 to 10.0 mmol/l MES, 50 mmol/l, (rest), pH 5,5-6,5.
This reagent is characterized by the accuracy of determining the activity of total α-amylase 100±5% reproducibility (coefficient of variation) of less than 5%. The shelf life of the reagent at 2-8°C is 2 years.
The inclusion of the new reagent components Triton X-100, which is a non-ionic surface-active agent, EDTA - forming complex compounds with most cations, as well as replacement of calcium chloride on the preservative calcium acetate can improve the stability of the reagent and extend its shelf life up to 2 years.
The determination of the activity of total α-amylase is carried out at a temperature of reagent 37°C as follows: 25 ál of sample add 1 ml of reagent and after 1 min start to read the optical density at equal intervals of time within 1-3 min at a wavelength of 405 nm, while the number of reads should be at least three. Actively the th total α-amylase (A) in u/l calculated according to the formula:
A=ΔΕ/min×3060, where ΔΕ/min - average change in optical density per minute; 3060 - the conversion factor by the equation the Bouguer-Lambert-Bera.
Determine significant differences of the proposed reagent, compared with the prototype are:
the reagent further comprises Triton X-100 and EDTA in experimentally selected, the optimal quantities, which improves the stability of the latter during storage;
in the reagent calcium chloride is replaced with a more suitable component is calcium acetate, which improves the stability of the reagent during storage;
liquid reagent ready to use, which can significantly simplify the procedure of analysis;
- all components in the reagent used in the optimal experimentally selected concentrations that can increase functionality and to extend the scope of the reagent.
The proposed reagent is clinically tested and approved for production, sale and use on the territory of the Russian Federation. The proposed reagent available for any clinical diagnostic, outpatient and biochemical laboratories and allows you to get accurate, reliable results of the analysis, the corresponding foreign standards.
To prepare the reagent doing weighed on an analytical balance for each component. The Perrin is placing them in glass beakers and dissolved in a small amount of distilled water. The solutions are thoroughly mixed on a magnetic stirrer. Solution components except GalG2CNP, transferred into a volumetric flask with a capacity of 1 liter and adjusted the pH under control of a pH meter to 5,73. Then add a portion GalG2CNP. The volume of solution was adjusted to the mark with distilled water and mix thoroughly. The result is a reagent for the determination of total α-amylase in serum, plasma and urine, containing the following components:
Sodium chloride, mmol/l to 50.0
KSCN, mmol/l - 400,0
Sodium azide, mmol/l to 90.0
EDTA, mmol/l 0,2
Calcium acetate, mmol/l 0,7
Triton X-100, vol.% - 0,075
GalG2CNP, mmol/l 0,5
MES, mmol/l to 50.0
Water - the rest.
When the pH value of the reagent is 6.0.
The reagent carried out analogously to example 1. Get reagent for the determination of total α-amylase in serum, plasma and urine, containing the following components:
Sodium chloride, mmol/l - 300,0
KSCN, mmol/l to 100.0
Sodium azide, mmol/l to 20.0
EDTA, mmol/l to 2.5
Calcium acetate, mmol/l 5,0
Triton X-100, vol.% - 0,075
GalG2CNP, mmol/l 10,0
MES, mmol/l to 50.0
Water - the rest.
When the pH value of the reagent is 5,5.
The reagent carried out analogously to example 1.
Get reagent for the determination of total α-amylase in serum, plasma, crowie urine, containing the following components:
Sodium chloride, mmol/l - 500,0
KSCN, mmol/l to 50.0
Sodium azide, mmol/l - 1,0
EDTA, mmol/l 5,0
Calcium acetate, mmol/l 10,0
Triton X-100, vol.% - 0,075
GalG2CNP, mmol/l to 20.0
MES, mmol/l to 50.0
Water - the rest.
When the pH value of the reagent is 6.5.
The determination of the activity of pancreatic α-amylase in the control sera.
To 1 ml of reagent, heated to a temperature of 37°C, was added 25 ál control sera "Precinorm U". After 1 min started to read the optical density at equal intervals of time within 1-3 min at a wavelength of 405 nm. The same procedure was repeated for the control sera "Precipath U", "c.f.a.s.", "Contornorm", "Contropath", "BioCal E". The results of the analysis are given in table 1.
|Control serum||Certified activity value of pancreatic α-amylase (IFCC), u/l||The result, u/l|
|"Precinorm U" (Roche, Germany)||40,8 (33,6-48,0)||40|
|"Precipath U" (Roche, Germany)||102 (84-120)||102|
|"c.f.a.s." (Roche, Germany)||162||164|
|"Contornorm" ("Biocon", Germany)||40,1 (32,9-47,3)||41|
|"Contropath" ("Biocon", Germany)||102 (84-120)||101|
|"BioCal E" ("Biocon", Germany)||171||177|
From table 1 it is evident that the proposed reagent provides high accuracy for determining the activity of total α-amylase in serum, plasma and urine.
The use of the reagent will allow for a comparison with prototype:
to increase the shelf life of the reagent up to 2 years by ensuring its stability;
- to expand the scope and improve the functionality of the reagent by providing opportunities to work with him in terms of clinical-diagnostic, outpatient and biochemical laboratories.
Reagent for determining the activity of total α-amylase kinetic colorimetric method containing the substrate 2-chloro-4-nitrophenyl-4-O-β-D-galactopyranosyl (GalG2CNP), sodium chloride, potassium radamisty, sodium azide, a water-soluble salt of calcium, 2-morpholinepropanesulfonic acid (MES) and water, characterized in that it complements the flax contains EDTA and Triton X-100,
and as the calcium salt is calcium acetate, in the following ratio of components:
|Sodium chloride, mmol/l||50,0-500,0|
|Potassium radamisty, mmol/l||50,0-400,0|
|Sodium azide, mmol/l||of 1.0 to 90.0|
|Calcium acetate, mmol/l||0,7-10,0|
|Triton X-100, vol.%||0,075|
|GalG2CNP, mmol/l||of 0.5 to 20.0|
if the pH of the reagent is 5,5-6,5.
SUBSTANCE: invention concerns a kit of reagents for pancreatic α-amylase activity test including a reagent 1 containing sodium chloride, potassium sulphocyanate, sodium azide, water-soluble calcium salt, monoclonal salivary α-amylase antibodies (MAB), 2-morpholinoethanesulfonic acid (MES) and water, and a reagent 2 containing 2-chloring-4-nitrophenyl-4-O-β-D-galactopyranosylmaltoside (GalG2CNP), MES and water, differing by the fact that the reagent 1 in addition contains EDTA and bovine serum albumin (BSA), and as calcium salt, it contains calcium acetate, and the reagent 2 in addition contains EDTA and sodium azide in the proportions specified in the patent claim.
EFFECT: enhanced stability of the set of reagents with maintaining high accuracy and result reproducibility of the analysis.
1 tbl, 4 ex
SUBSTANCE: invention relates to field of medicine, namely to methods of predicting post-operational complications, namely to methods of predicting development of scars after previous acne. In order to predict scar development content of receptor antagonist of interleukin-1 (RAIL) is determined during 15 days after resolution of inflammatory process. Scarless development of process is diagnosed at level RAIL in peripheral blood serum after disease being within physiological norm (300-800 pg/ml). If level of RAIL is lower than said norm prediction of acne complication in form of skin scars is diagnosed. Possibility of development of hypertrophic scars is predicted if RAIL concentration is lower than 200 pg/ml.
EFFECT: method makes it possible to predict type of complications after previous acne, therefore correcting therapy carried out in due time can prevent risk of skin scar formation and improve patient's life quality.
2 cl, 1 tbl, 4 ex
SUBSTANCE: developing hypoxia in a pregnant woman of the third trimester of gestation is predicted by evaluating peripheral blood oxyhemoglobin (HbO2) and 2,3 diphosphoglycerate phosphatase (2,3DPG) concentrations. A discriminator (D) is calculated by formula D=+17.072·2.3DPG + (-0.041·HbO2), 2,3DPG - concentration, mol/l, HbO2 - amount, %. The D value within 97.09-112.37 enables to predict threatened hypoxia in a pregnant woman had an acute condition of herpes virus infection.
EFFECT: use of the method allows well-timed detection of a risk group and correct prediction of developing hypoxia.
SUBSTANCE: threatened reduced erythrocyte oxygenation in a pregnant woman suffering an acute attack of bronchial asthma in the first trimester of gestation is predicted by evaluating oxyhemoglobin (HbO2) and 2,3 DPG (2,3 diphosphoglycerate phosphatase). This is followed by calculating a D discriminator value by formula D=18.05·2,3DPG + (-0.075·HbO2), and observing the D value within 88.66 to 102.42, threatened reduced erythrocyte oxygenation that leads to hypoxia is predicted.
EFFECT: use of the method enables predicting threatened reduced erythrocyte oxygenation in a pregnant woman with the acute attack of bronchial asthma.
SUBSTANCE: differentiated detection large and small circulating immune complexes in blood serum is ensured by a follow-on examination of blood serum clarified by short centrifugation with a method of circulating immune complexes precipitation PEG -6000 of the end concentration 4 % and 6 %. Thereafter, the results are recorded by a turbidimetric method with using a microplate spectrophotometer in a two-wave mode: basic - 340 nm, reference filter - 620 nm. Duration of an incubation step at temperature +18-25°C is 15 minutes. Using the method enables higher effectiveness and reliability of determining the level of large circulating immune complexes, results reproducibility, as well as differential measurement of the level of small CIC which are a diagnostically significant indicator of human body immune responsiveness in many types of a pathology, decreased volume of analysed serums to 0.06 ml, and incubation duration to 15 minutes.
EFFECT: method is suitable for clinical screenings.
1 dwg, 1 tbl, 1 ex
FIELD: measurement equipment.
SUBSTANCE: invention refers to biophysics. In order to determine calcium concentration on the basis of discharged photoproteins, bioluminescent reaction of photoproteins and calcium ions is performed, intensity of the solution fluorescence is measured, calibration dependence of fluorescence intensity on calcium concentration in double logarithmic coordinates is built and logarithm value of calcium concentration is determined as per the logarithm of the measured fluorescence intensity. Fluorescence is initiated with the light source after bioluminescent reaction is completed. Fluorescence intensity is measured at the specified length of excitation and recording wave.
EFFECT: method allows determining calcium concentration in various media as per fluorescence intensity of discharged photoproteins, which allows performing continuous measurements of calcium concentration in "in vivo" system.
2 dwg, 6 ex
SUBSTANCE: when admitted, an acuity patient is analysed for total protein concentration and creatine phosphokinase activity in blood serum. If the analysed values are within normal limits, the absence of skeletal muscles injures is diagnosed; total protein concentration being within normal limits and creatine phosphokinase hyperactivity indicate the presence of an accompanying injury of the skeletal muscles, while lowered total protein concentration with underlying creatine phosphokinase hyperactivity shows progressing hypoproteinemia.
EFFECT: method provides pre-clinical detection of the presence of the accompanying injures of the muscles and progressing hypoproteinemia.
SUBSTANCE: invention relates to field of medicine, in particular toxicology and resuscitation science and can be used for early prediction of pneumonia development in patients. On the first day of staying in hospital functional state of albumin in blood serum is analysed in patients by fluorescent method. Intensity of K-35 probe fluorescence in blood serum albumin in medium with high ionic power is determined. If value of K-35 probe fluorescence intensity in albumin is lower than or equals 36 conv.units, development of pneumonia in patient is predicted.
EFFECT: method allows to increase efficiency of performed treatment in said category of patients.
2 tbl, 4 ex
SUBSTANCE: invention relates to medicine, namely to internal diseases, diagnostics. Method is based on determination of general oxidant activity (GOA) and general antioxidant activity (GAA) with further determination of oxidant index (OI), which equals ratio of GOA to GAA. In accordance with the invention oxidant activity is determined by degree of echinochrome A oxidation by components of oxidant system of blood serum or plasma, and general antioxidant activity is determined by degree of echinochrome A oxidation with chloramine B, added to blood serum or plasma. OI index of healthy donor is taken as a unit. OI value higher than a unit testifies to disbalance of general oxidant status of organism. Method ensures up to 20 fold reduction of amount of analysed serum or plasma in comparison with standard methodology, reduction for carrying out analysis from 72 hours to 2 hours. When ranging it is possible to perform analysis of more than 500 samples during one day, using microboards and microplate spectrophotometers.
EFFECT: method is simple in implementation, economical and does not require large volume of thermostatically controlled chambers.
1 tbl, 2 ex
SUBSTANCE: invention relates to medicine, in particular to physiology, biochemistry and can be used for express diagnostics of organism adaptation to hypokinesia both in usual conditions, and during space flights. Blood parametres are determined in patients only with hypokinesia and with hypokinesia at the background of desmopressin introduction, obtained results are compared and conclusion is made, as parametres mass-spectral determination of proteomic profiles of blood serum in mass range from 1000 to 17000 with processing of samples with magnetic particles MB WCX and MB IMAC Cu is carried out, and if there is no change in signal intensity after introduction of desmopressin with pre-fractioning by magnetic particles MB IMAC Cu m/z=9133 in comparison with initial data, as well as with reliable reduction of peak intensity m/z=1297, m/z=2769, m/z=7764, m/z=6432, m/z-6630 when pre-fractioning with magnetic particles MB WCX in these users stage of organism adaptation to hypokinesia with introduction of desmopressin is diagnosed.
EFFECT: method acceleration, possibility to determine about 20 parametres during one analysis, minimal amount of biomaterial (5-10 mcl), as well as possibility of said technique automation.
5 cl, 3 tbl, 1 dwg
FIELD: medicine, analytical biochemistry.
SUBSTANCE: invention relates to laboratory methods of investigations. Method involves sampling specimen from patient to be inspected, extraction of serotonin and histamine from a specimen, chromatography of extract and determination of concentration of serotonin and histamine by the fluorescence intensity value. Saliva is used as biological fluid. Saliva by volume 1 ml is extracted with 4 ml of 1 N hydrochloric acid solution, 2 g of anhydrous potassium carbonate and 5 ml of mixture of butanol and chloroform in the ratio 3:2 are added, extract is shaken up and centrifuged. Organic phase (4 ml) is sucked off from extract and passed through chromatography column (diameter is 3 mm, height is 16 mm) filled with ion-exchange resin KB-4 or KB-4P-2 or Bio Rex-70 in H+-form, size of granules is 0.1 ± 0.02 mm. Histamine is eluted with 4 ml of 0.1 N hydrochloric acid at the rate of eluting solution 0.4 ml/min. Histamine concentration is determined by reaction with ortho-phthalic aldehyde dissolved in ethanol. Serotonin concentration is determined by reaction with ninhydrin in organic passed through column. Method provides assaying the saliva concentration of serotonin and histamine with high precision.
EFFECT: improved assay method.
SUBSTANCE: method involves studying blood samples with venous blood mixed with vital stain like methylene blue. Degree of vital stain absorption by erythrocytes is determined by applying photocolorimetry. The value drop being more than 25%, extracorporal detoxication is to be predicted as ineffective.
EFFECT: simplified method.
FIELD: medicine, infectology.
SUBSTANCE: one should detect the level of terminal stable metabolites of nitrogen oxide (NOx) in whole blood. At its value ranged 39.6-86.0 mcM/l one should evaluate hemorrhagic fever accompanied with renal syndrome (HFRS) of average severity, at NOx value ranged 86.7-141.5 mcM/l - severe form of HFRS and at its values ranged 88.2-128.6 mcM/l at the background of pronounced clinical picture - as complicated disease flow. The method enables to shorten the terms for carrying out the assays.
EFFECT: higher accuracy of evaluation.
3 ex, 1 tbl
FIELD: medicine, biochemistry.
SUBSTANCE: in blood serum one should detect the level of lactoferrin and biliary acids. At their ratio being equal to 5-17 it is necessary to detect chronic hepatitis of high activity.
EFFECT: higher accuracy of detection.
FIELD: medicine, dermatology, clinical laboratory diagnostics.
SUBSTANCE: the present method deals with detecting the focus of neutrophilic phagocytic activity lesion in capillary blood. At the values of cells' capacity to phagocytosis in percentage and phagocytic number on the 10th d of therapy being below 20% and 3.3, correspondingly one should evaluate therapeutic efficiency to be low, if it is above 40% and 4.0, correspondingly - as high.
EFFECT: higher accuracy of evaluation.
2 ex, 3 tbl
SUBSTANCE: method involves studying blood serum, processing obtained data and setting disease diagnosis. The study is carried out by preparing dried blood serum sample as suspension in Vaseline oil and doing the infrared spectroscopy analysis in the bandwidth of 120-1000 cm-1 and determining absorption strip peak heights having maximum at 1180; 1165; 1160; 1150; 1130; 1070; 1025 cm-1 and then calculating the following two ratio groups, the first of which is ratio of peak height with maximum at 1165 cm-1 to 1150 cm-1; 1160 cm-1 to 1130 cm-1; 1070 cm-1; 1025 cm-1. The second group has ratio peak having maximum at 1165 cm-1 to 1160 cm-1; 1180 cm-1 to 1130 cm-1; 1065 cm-1; 1070 cm-1. The obtained three-dimensional distribution of the first group is projected to frontal plane for calculating two-dimensional coordinates and comparing to flat reference diagnostic images of hepatic pathologies and to a normal reference diagnostic image represented as flat polygons which boundaries are given by the following values. The norm is represented by X(-2.3;2.0;4.0;4.0) and Y(1.6;0.8;0.8;1.6), respectively. Oncology is represented by X(1.7;1.7;0.0;0.0) and Y(1.9;1.25; 1.25;1.9). Hepatites are represented by X(1.9;2.2;1.8;1.4 and 1.9;1.8;4.0) and Y(1.9;1.9; 0.5;0.5 and 08;0.5;0.8). Cirrhosis is represented by X(1.9;2.6;1.4) and Y(1.6;0.8;0.4). Diseases are differentiated by interpreting point position within particular area. Three-dimensional distribution of the second group is projected to frontal plane and compared to diagnosis images of pathology and norm. Coordinate values of the second group are as follows: norm - X(1.8;2.9;2.5;1.5), Y(2.7;2.0;1.2;1.6); oncological cases - X(0.27;0.67;0.63), Y(0.27;0.67;0.3); hepatitis - X(1.5;2.5;2.4;1.2), Y(1.6;1.2;0.2;0.9); cirrhosis - X(1.1;0.9;0.9). Final diagnosis of pathology is set when particular data values belong to the corresponding pathology zone in both cases.
EFFECT: high accuracy of diagnosis.
SUBSTANCE: method involves studying biological material by applying infrared spectroscopy techniques. The obtained data are processed and diagnosis is set. Blood serum is used as the biological material. The study is carried out by preparing dried blood serum sample as suspension in Vaseline oil and doing the infrared spectroscopy analysis in the bandwidth of 120-1000 cm-1 and determining absorption strip peak heights having maximum at 1170; 1165; 1160; 1150; 1140; 1060; 1050; 1040; 1025 and then calculating the following ratio values like peak height with maximum at 1160 cm-1 to 1140 cm-1; 1165 cm-1 to 1150 cm-1; 1040 cm-1 to 1025 cm-1. The obtained distribution of this group is projected to frontal plane for calculating two-dimensional coordinates and comparing to flat reference diagnostic images of prostate pathologies and to a normal reference diagnostic image represented as flat polygons which boundaries are given by the following values. The norm is represented by X(-1.15;-0.9;0.45;0.0;-0.65) and Y(0.99;4.2;0.9;0.46), respectively. Pathology by X(-1.15;-1.15;0.35;0.0;0.65) and Y(0.99;-0.03; 0.48;0.09;0.46). The norm and pathology are differentiated. Additional mathematical processing is carried out on infrared spectra of blood serum samples of patients belonging to pathology image according to parameter values. First of all, three-dimensional distribution is calculated as peak having maximum at 1160 cm-1 to one having maximum at 1150 cm-1; 1170; 1160 cm-1; 1160 cm-1 to 1025 cm-1. It is projected then to frontal plane and compared to diagnosis images of prostate adenoma and images of prostate carcinoma. The second group relationships the following values are used: oncological cases - X(0.28;0.77;1.24;0.96), Y(0.75;0.46;-0.13;-0.02); adenoma - X(0.28;1.24;2.21;1.24;0.77), Y(0.75;1.24;-0.12;-0.13;0.46). Differential diagnosis of pathologies is set by interpreting point position within particular pathology image.
EFFECT: high accuracy of differential diagnosis.
SUBSTANCE: method involves determining mean cytochemical coefficient of lipid accumulation in peripheral blood leukocytes in conditional units before beginning therapy application (MCC1) and in 2-3 or 5-6, or 10-12, or 20-24 months of therapy application (MCC2). Therapy effectiveness coefficient is calculated in conditional units from formula K= MCC2/MCC1. The value being equal to or greater than 1, leprosy therapy is predicted to be effective.
EFFECT: simplified prognosis method.
1 dwg, 1 tbl
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: method involves pouring venous blood treated with heparin into five conic test-tubes in the amount of 0.1 ml. The first three of them contain 0.1 ml of non-colored latex suspension with particle size of 1.5 mcm, the fourth one contains 0.1 ml of medium 199 and 0.1 ml of 0.1% aqueous solution of tetrazole nitro blue, the fifth one contains .1 ml of latex suspension and 0.1 ml of 0.1% aqueous solution of tetrazole nitro blue. The first test-tube is incubated in thermostat for 5 min at37°C, the second one for 30 min, the third one for 1 h, the fourth and the fifth one for 40 min. Smears are prepared from 0.2 ml of incubation mixture on glasses and dried at 37°C, fixed in burner flame, stained with 0.1% aqueous solution of tetrazole nitro blue, repeatedly dried and studied with microscope under immersion with magnification of 90x10. Test results are evaluated from absorption activity in phagocytosis reactions in determining the number of phagocytes, phagocytic number, phagocytic integral index and phagocytosis rate values. Tetrazole nitro blue test response is determined by counting formazan-positive cell number, calculating cytochemical activity index and tetrazole nitro blue test stimulation index.
EFFECT: accelerated test; high accuracy and low cost of examination.
1 dwg, 3 tbl