RU2428696C2 - Specifying diagnostic technique for acute exacerbation of chronic obstructive pulmonary disease - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: specifying diagnostic technique for acute exacerbation of chronic obstructive pulmonary disease is implemented by blood examination. The blood values are estimated by granulocyte binding lymphocyte cytoleukogram with calculating a resetting index (RI) as a relation of LSG-1 (lymphocyte +1 granulocyte) to an amount of loose-lying lymphocytes, LSG-2 (lymphocyte +2 granulocytes) to an amount of loose-lying leukocytes. If the RI value is more than 0.8 and the RI LSG-2 value is reduced less than 0.11, acute exacerbation of chronic obstructive pulmonary disease is diagnosed in the patients.

EFFECT: use of the declared method allows accelerating and providing more precise diagnostics of chronic obstructive pulmonary disease.

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Description

The invention relates to medicine, namely to pulmonology, and can be used to refine the diagnosis of exacerbation of chronic obstructive pulmonary disease (COPD).

COPD is one of the leading causes of morbidity and mortality in the modern world. Mortality from COPD takes the 4th place among all causes of death in the general population, which is about 4% in the structure of total mortality. A feature of the course of COPD, like any other chronic pathology, are exacerbations, each of which aggravates the progression of COPD, thereby accelerating and aggravating the development of DN.

The criteria for exacerbation of COPD are clinical symptoms (increased severity of dyspnea and cough, an increase in the number of wheezing, an increase in sputum production and an increase in its purulence, the appearance of congestion in the chest, the appearance of peripheral edema), a decrease in the level of high pressure and blood gases, and a decrease in exercise tolerance. arising suddenly and lasting at least 2 days.

However, quite often, especially with severe exacerbation of COPD, there is not an increase in sputum production, but, on the contrary, its delay, which makes it impossible to assess the degree of its purulence. Exacerbations can begin gradually, with unexpressed clinical symptoms, which complicates the timely recognition of exacerbations. In addition, most patients with COPD have a combined extrapulmonary pathology, most often cardiovascular diseases, which are often decompensated against exacerbations of COPD, which, in turn, worsens the prognosis and complicates the management of this category of patients.

Therefore, timely diagnosis, relief and prevention of exacerbations come to the fore in the management of a patient with COPD.

The closest analogue is the method proposed in the application No. 97114285 with the date of receipt of 05.08.1997, by which the patent for the invention No. 2178176 of 01.10.2002 was received.

The purpose of the invention is to accelerate and improve the accuracy of the method of clarifying diagnostics of exacerbation of COPD.

The method is based on the ability of lymphocytes and neutrophils to form autologous rosettes in vitro.

The method is as follows.

The subject takes blood from a vein (3 ml) into a test tube with heparin, 25 units of heparin per 1 ml of blood. Blood is diluted with a Hanks solution in a ratio of 1: 1. At the bottom of the dry tube, 3 ml of density gradient is poured p = 1.077 g / ml. To control viability, cells were stained with trypan blue. The percentage of stained cells did not exceed 3-4. Layered diluted blood on a density gradient, preventing mixing media. Centrifuge 40 minutes at 1500 rpm. After centrifugation, remove the supernatant with a pipette and collect the upper ring, consisting of lymphocytes, into a dry tube. The lower plasma layer containing granulocytes is collected in another tube. Hanks solution is added to 5 ml, resuspended and washed at 1500 rpm for 10 minutes. Remove the supernatant to 0.1 ml. Hanks solution is added to obtain a suspension of lymphocytes in a volume of 1 ml, granulocytes - 0.7 ml. Resuspend. Cells are counted in the Goryaev chamber and adjusted to the desired concentration (lymphocytes - 4.5 × 10 ⁹ / L, granulocytes - 3.0 × 10 ⁹ / L). Transfer lymphocytes to the backprint, granulocytes on a coverslip and incubate for 1 hour at 37 ° C in an thermostat. After incubation, the supernatant is removed from the granulocytes and lymphocytes are added to the granulocytes. Incubated at a temperature of 37 ° C for 1 hour in a thermostat. Then add 1 drop of glutaraldehyde 3% and incubated for 5 minutes at room temperature. Remove the supernatant. Smears are dried. Stained by May-Grunwald. Dry at room temperature. Using a light microscope, determine the number of free-lying lymphocytes and rosette-forming. To increase the accuracy of the results, counting is performed on 300 cells in 3 fields of view with the further construction of a cytolucogram.

The analysis of the cytoleukogram is carried out as follows. The percentage of free-lying lymphocytes and rosette-forming ones is estimated as lymphocyte +1 granulocyte (LSH-1), lymphocyte +2 granulocyte (LSH-2), lymphocyte +3 granulocyte. The index of rosette formation (RTI) is calculated - the ratio of the values between LSH-1, LSH-2, LSH-3 and free-lying lymphocytes and with an increase in the RTI of LSH-1 more than 0.8, a decrease in the RTI of LSH-2 less than 0.11, patients are diagnosed with an exacerbation of chronic obstructive pulmonary disease.

We have studied the features of the LSH cytoleukogram in patients with COPD (without severity) in a stable course (group 1) and during exacerbation (group 2). By age, gender, duration of the disease, the smoker index of the group were equivalent. The diagnosis was made on the basis of clinical and instrumental examination methods, in accordance with the standards for the diagnosis and treatment of patients with COPD (2006).

The results obtained, characterizing the features of the LSH cytoleukogram in patients with COPD with a stable course and during exacerbation are presented in table 1.

As can be seen from the table, in patients with COPD during exacerbation, in contrast to the group of patients with a stable course, the indicators of LSH-1 increase significantly (p <0.001), indicating an inflammatory process and significantly (p <0.001), the parameters of LSH-2 decrease, indicating for the presence of immunodeficiency, the IRO of LSH-1 significantly increases and the IRO of LSH-2 decreases. No significant differences in the content of LSH-3 were detected.

The developed diagnostic method allows you to obtain reliable data within 4 hours. The method does not require special sterility, scarce reagents, expensive devices and is simple to perform, economical and has no contraindications.

Table 1 Indicators of granulocyte-binding lymphocytes in patients with COPD with a stable course and with exacerbation Group P Granulocyte binding lymphocytes (M ± m)% SVL LSG-1 LSG-2 LSG-3 1 group 23 56.48 ± 0.55 30.46 ± 0.34 9.47 ± 0.40 0.63 ± 0.75 IRO 0.5 0.16 0.01 2 group 25 53.25 ± 0.55 ** 43.0 ± 0.66 **** 6.12 ± 0.25 *** 0.60 ± 0.45▪ IRO 0.8 ** 0.11 ** 0.01 Note: ▪ * - a confidence indicator between the COPD group of moderate course with a stable course and the COPD group during exacerbation and the control group; • p> 0.05; ** p <0.01, *** p <0.001 SVL - free-lying lymphocytes; LSH-1 - lymphocyte +1 granulocyte), LSH-2 - (lymphocyte +2 granulocyte), LSH-3 - (lymphocyte +3 granulocyte); IRO - Rosette Index, COPD - Chronic Obstructive Pulmonary Disease

Information sources

1. Emelyanov A.V. Diagnosis and treatment of exacerbations of chronic obstructive pulmonary disease // Russian Medical Journal. - 2004. - T. 13, No. 4. - S.183-189.

2. Chuchalin A.G. White paper. Pulmonology. M., 2003 .-- P.67.

3. Georgupolos D, Antonisen NR. Symptoms and signs of COPD. In: Chemiak NS, ed. Chronic obstructive pulmonary disease. Philadelphia: Saunders, 1991 .-- P.357-363.

4. Standards for the treatment and diagnosis of chronic obstructive pulmonary disease: Per. from English under the editorship of A.G. Chuchalina M., 2004.

5. Ovcharenko S.I., Leshchenko I.V. Current problems in the diagnosis of chronic obstructive pulmonary disease. Russian medical journal. 2003. - No. 4. - T. 11. - S. 160-164.

6. Respiratory medicine. Manual edited by Acad. RAMS A.G. Chuchalina, Volume 1. M., “GZOTAR-Media”, 2007. - P.616-632.

Claims (1)

A method for improving the diagnosis of exacerbation of chronic obstructive pulmonary disease by examining a blood, characterized in that the blood counts are estimated by a cytoleukogram of granulocyte-binding lymphocytes with further calculation of rosette formation indices (RTI) as the ratio between LSH-1 (lymphocyte + 1 granulocyte) to the number of free-lying lymphocytes, LSH- 2 (lymphocyte +2 granulocytes) to the number of free-lying white blood cells, and with IRO LSH-1 more than 0.8, a decrease in IRO LSH-2 less than 0.11, patients diagnosed with exacerbation of chronic uktivnoy lung disease.