SU1617382A1 - Method of differential diagnosis of tuberculosis and acute pneumonia - Google Patents

Abstract

This invention relates to medicine and can be used in the differential diagnosis of tuberculosis and acute pneumonia. The aim of the invention is to improve the accuracy and acceleration of the method. To do this, erythrocytes are isolated from 2 ml of venous blood, erythrocyte shadows are obtained by hemolysis, followed by washing the membranes from hemoglobin, and the content of TBA-active products in the erythrocyte membranes is determined. In another portion of erythrocyte shadows, superoxide dismutase (SOD) activity is determined. To do this, ethanol-chloroform-water is added to the shadows, shaken and settled before the phases are separated. Then, superoxide dismutase activity in inhibiting the reduction of NST is determined in the water-alcohol phase. With the values of TBA-active products of lipid peroxidation (POL) 8.9-9.5 nM per 1 mg of protein and SOD activity 171-174 U, tuberculosis is diagnosed, and with values of POL products 7.0-7.5 nM and SOD activity 179-180 U - acute pneumonia. The use of the invention makes it possible to establish the diagnosis earlier and to increase the accuracy of the differential diagnosis up to 92%.

Description

The sediment is washed 3 times and the sedimentation of erythrocyte membranes is opaque.

Gh,:.

1 color used in the experiments.

: The suspension of erythrocyte membranes is incubated in a standard mixture consisting of Oj3 ml 12 X Mora salt, 0.3 ml 2 X pyrophosphate and 0, .3 ml 1 W NADPH2. I start the reaction by adding 0, t mp membrane suspension

; red blood cells. Incubation is carried out in a water bath at t 37 ° С s

: 20 minutes. The reaction is stopped by adding: T ml of 30% trichloroacetic acid (THU). Sediment is removed centered

 by fugging for 10 min at 6000 rpm. To develop the color, the tubes with a mixture of supernatant and thiobarbituric acid (TBC) are placed in a water bath at t and incubated for 10 min. From: optical density is measured at a spectrophotometer at 535 nm. The peroxidation rate is expressed in nan moles per 1 mg of protein.

To determine the activity of superoxide dismutase (SOD) in erythrocytes, erythrocytes were isolated from 1 ml of heparinated venous blood by centrifugation for 5 min at lOOO rpm

 at t, the plasma is discarded, the erythrocytes are flushed with 10 ml of cold saline, separated by repeated centrifuging at 3000 rpm for 5 minutes. Hemo-L11; h is carried out by adding 5 ml of di: styled water, followed by agitation for 5 minutes; a dibasic potassium phosphate is added to the supernatant, ethanol: chloroform is mixed with water, mixed vigorously: and left to separate into two phases: upper - water-alcohol and lower - chloroform. The aqueous-alcoholic layer has a distinct yellow-green dye caused by erythrococprecrein exhibiting superoxide-dismutase activity. In the resulting preparation, it is determined

; activity according to method i based on measuring the degree of inhibition of the staining rate of blue nitrotetrazol. The reaction mixture volume. , 5 ml contains, µmol: pyrophosphate mixture 16; EDTA 7, nitrotetraz-rly blue, 50, phenazine metasulfate 4,5, NADPH 50 and the extract with the test

. an enzyme. Per unit of SOD activity take the amount of enzyme

which inhibits the degree of discoloration of nitrotetrazol blue from 50%. SOD enzyme activity is expressed in units of 1 ml of blood.

Example 1, Patient A, on admission, noted febrish, slight shortness of breath at rest, frequent inflammatory diseases of the respiratory tract in the past, physical examination of the lungs showed dullness of percussion sound, sonorous wet and dry rales, the X-ray process had a root lobe, root structure lung was changed. Mantoux test - 6 mm. Microbacteria turebculosis were not detected. Immunological examination; the level of E-ROCK was 19.2%, the percentage of blast infection in RBTL for phytohemagglutinin 26%, for tuberculin 1.3%. In a special biochemical study, it was established that the FLOOR level is 7.2 nM, the activity of SOD is 179.1 U (within the normal range). On the basis of clinical, radiological, biochemical, and immunological indications, acute pneumonia has been diagnosed. Twenty days after trial antibacterial therapy with broad-spectrum drugs, complete resorption of infiltrative changes was observed,

Example 2, Free T, on admission was noted subfebrile, cough, weakness. During physical examination, dullness of percussion sound, single moist rales, the process had an upper lobe location in S, S. The Mantoux reaction was 10 11 U1M. In the immunological study: the level of E-ROCK is 33%, the percentage of blasts on the RBTL for phytohemagglutinin is 35%, for tuberculin is 1j7%. The LPO level was 9.1 nM, SOD activity was 172.5 U (below normal). Based on clinical, radiological and laboratory; data was diagnosed with infiltrative pulmonary tuberculosis. Anti-tuberculosis therapy was prescribed, on the background of which improvement came, and the incidence of intoxication was reduced. The growth of microbes of tubsfculosis after May 2 showed positive results.

Note 3, Patient D., 28 years old The condition of the patient upon admission to the clinic is moderate. Complaints about a cough with a small amount of mo1

Moles, weakness, sweating, temperature rise to STjS C. Physical examination of the lungs showed some weakening of breathing, single moist rales. Reaction to a Mantoux test with 2 TE - 6 mm papule. In the sputum by sowing, microbacteria were not found. X-ray; polymorphic sleepers in the region of the upper lobe of the right lung. To conduct differential diagnostics, additional biochemical studies were performed: the POL level was 8.9 nM, TOD activity was 172.0 U. Based on clinical and radiological studies, as well as data from a special biochemical study on the 5th day of admission, the diagnosis of infiltrative tuberculosis of the upper lobe of the right lung was diagnosed. Conducted anti-tuberculosis treatment.

PRI me R 4. Patient G., 46 years old. The patient's condition at admission to the clinic was satisfactory, the temperature was within the normal range. Complaints of dry cough. During the physical examination of the lungs against the background of weakening of the body, single moist rales were heard. Reactions to the Mantoux test with 2 TE 10 mm. In the sputum by the method of sowing tuberculosis microbes were not found. Radiographically: heterogeneous darkening of the upper lobe of the right lung,

A detailed biochemical study revealed an increase in the activity of POL to 9.5. Hl-1 and reduction of SOD activity to 173.0 U.

On the basis of a clinical X-ray study, as well as additional biochemical studies on the 3rd day, a diagnosis was made: infiltrative tuberculosis of the upper lobe of the right lung.

PRI me R 5 Patient A., 3 years. The condition is medium yellow. Zhajioby cough with sputum, alone at rest, temperature rise to 38 ,. Physical examination of the lungs showed dullness of percussion sound, dry moist rales. Reactant Mantoux test with 2 TE - nanyns V mm. Radiographically: homogeneous darkening of the upper lobe of the right lung, changing the structure of the root of the lung. When, a special biochemical research institute has been established: the POL level is equal to 7.2 Hl-1, which is significantly lower than 7382-6

nor, obtained from a patient with tuberculosis, the activity is 179.1 U (within the normal range).

Diagnosis; acute pneumonia (krupnozna).

Example 6. Ill G., 45 years old. The condition of the patient is moderate. - Complaints of cough with sputum, once. 10 At a fnzikalny research gpitucheniye of a percussion sound, moist rales is noted. The reaction to the Mantoux test is negative.

Radiographically: homogenous darkening Sj of the right lung. With

a special biochemical study showed an increase in the POL level to 7.2 nM, SOD activity equal to 179.0 U, Diagnosis: acute pneumonitis,

The material was subjected to statistical processing by the method of variation statistics using Student's criteria. On the basis of statistical processing, the boundaries of the values of the POL level and SOD activity were chosen. In pneumonia, the level of FLOOR is 7.3 ± 0.14,, 00.1, SOD activity .179.5 ± 0.1, P 0.05, Pr1g tuberculosis 0 POLO 9.2 + 0.16, ..001,, SOD 172.5 ± О, 3, Р с 0.05. Control values (donors) FLOOR 4,3 ± 0,18, SOD 130,0 t 0,59, where P is the reliability compared with the control.

The values of these indicators were analyzed in 79 patients with tuberculosis and in 67 patients with acute pneumonia. According to this data, the frequency of positive cases, i.e. the coincidence of the diagnosis, Q delivered by the proposed method, with the final diagnosis is 92 + W; the frequency of negative cases is 5 + 2%, the percentage of false positives is 3 + 2%. The accuracy of prototype 5 is 63.5%.

The use of the proposed method makes it possible to establish the diagnosis of the disease at an early stage with high accuracy and thereby initiate a proper and temporary treatment. The method is simple in execution.

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Claims (1)

Invention Formula Differential diagnosis of tuberculosis and acute pneumonia by means of a biochemical examination of the patient's blood, characterized in that, in order to improve the accuracy of method and its acceleration, in the membrane and activity of superoxide dismutase In the wounds of erythrocytes, coderum-171.0-174.0 U was diagnosed to diagnose tuberculation of TBA-active products, and at values of lipid peroxidation products and lipid acid oxidation, 7.0-7.5 nM superoxide dismutase and when the value of superoxide dismutase is 119.0-180.0 U. TBA-active products 8.9-9.5 nAmid tnevmoniya.