SU1675728A1 - Method of atopic bronchial asthma diagnosis - Google Patents

Abstract

The invention relates to medicine, namely to allergology. The goal is to improve the accuracy of the method. The goal is achieved by additionally determining the content of cortisol or thyroid stimulating hormone in the blood. The diagnosis of the disease is carried out according to the system of the location of these points in the coordinate system. The method allows differentiation of atonic bronchial asthma from nonspecific lung diseases.

Description

The invention relates to medicine, in particular to allergology, and can be used to diagnose atonic (non-infectious-allergic) bronchial asthma, not complicated by bronchopulmonary infection, and infectious-dependent nonspecific bronchopulmonary diseases, including chronic non-allergic bronchitis.

The aim of the invention is to improve the accuracy of the method due to the differential diagnosis of nonspecific diseases.

The method is carried out as follows.

Patients with a bronchospatic syndrome in history are examined during clinical remission. Take 2-3 ml of venous blood in a tube with an anticoagulant and 0.3 ml of a dry tube to obtain serum. Mononuclear cells are isolated from blood in a density gradient and washed in a conventional manner.

cell suspension in medium 199 with a concentration of (10-12) 110 cells / ml, stained (final dye concentration 25 µm); mix and expose at room temperature for 5 minutes. Then a drop of the suspension is applied onto a glass slide, covered with a cover glass, a drop of immersion oil is applied on top and placed under the microscope objective. The measurement of the fluorescence intensity of the dye in lymphocytes is carried out on a fluorescent microscope (for example, the LUMAM series) with a photometric cap. The maximum of fluorescence excitation spectrum in lymphocytes is 400-420 nm, fluorescence - 520-540 nm. The fluorescence intensity of 50-100 lymphocytes is measured, and the average amount of luminescence for this patient is calculated.

In the blood serum of patients, the concentration of cortisol or thyroid-stimulated hormone or both are determined in parallel using a radio-immune method.

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In the coordinate system, the values of the measured parameters are laid, the point of intersection on the plane is plotted. If this point lies below the previously designated border of the forms of bronchospastic syndrome, then atonic bronchial asthma is diagnosed, and if the point lies above the border, then it is a nonspecific disease.

The boundary of separation of the forms of bronchospasm syndrome in the specified coordinate system is established before the start of application of the method and is used in all subsequent diagnostics with constant operation of the instruments.

PRI me R 1. Mononuclear cells were isolated from heparinized (10 units / ml) venous blood of a patient by centrifugation in a Ficoll-verografin density gradient at 400 g for 40 minutes, then washed three times in saline with phosphate buffer (pH 7.4) and once in medium 199 at 200 g for 5-7 minutes. Cells are resuspended in medium 199 to the desired concentration. In a glass test tube, to a 0.1 ml suspension is added with a microsyringe 1 µl of a 2.5 mg 1 solution of dye in dimethylformamide.

Measurement of the intensity of its fluorescence in lymphocytes is carried out using a luminescent microscope. The object is illuminated from above through an illuminator and an opaque.

As a standard for luminescence, a reference fluorescent plate of ZhS 19 uranium glass with a thickness of 0.03 mm, included in the kit, is used. The reference plate is placed under the lens.

In serum, the concentration of cortisol or thyroid stimulating hormone is determined by radioimmuno method. Using a calibration curve based on the radioactivity of the standards, the values of the concentration

concentration of hormones in the studied samples. For a korszola, the boundary passes through the axis of ordinates at a point of 25 mV. and at an angle of 11 ° to the horizontal axis, for thyroid stimulating hormone - at a point of 41 mV, at an angle of 3 °.

Clinical diagnosis: bronchial asthma of infectious genesis of the 1 stage of mild course, remission phase, sensitization to house dust allergen.

Example 2, Patient, 22 g, complaints of attacks of suffocation, difficulty breathing out, cough with a small amount of sputum, which is difficult to expel. More than 2 g, clinical and anamnestic data did not allow to specify the diagnosis. In the study of respiratory function, a positive test with bronchodilator was detected,

When measuring the dye luminescence in lymphocytes is 61.4 mV, serum cortisol concentration is 540 nM / l, hormone is 1.6 mIU / l, the intersection points of these coordinates on the planes for the corresponding hormones are located above the accepted division lines, therefore, in a patient suggest the presence of non-specific disease.

Thus, the proposed method allows detection of chronic bronchial infection in a patient with atopic bronchial asthma, which significantly influences the further treatment tactics.

Claims (1)

The invention of a method for diagnosing atonic bronchial asthma by fluorescent analysis of blood lymphocytes stained with 3-methoxybenzathron, characterized in that, to increase the accuracy of the method, the content of cortisol or thyroid stimulating hormone is additionally determined in blood and at cortisol values of 50011.7 nM / l or thyroid-stimulating hormone 2.5 ± 0.15 mIU / l diagnoses the disease.