SU1359743A1 - Method of determining sensitization of leucocytes - Google Patents

Abstract

The invention relates to an immunology. The purpose of the invention is to reduce the sensitivity of the method. Blood samples with the test allergen are incubated. Then blood smears are prepared and stained with them. The content of modified leukocytes in the test and control smears is counted and the leukocyte sensitization index is determined. The method allows to diagnose allergic processes associated with infectious, non-infectious and autogenic allergens. Once ate 00

Description

This invention relates to medicine, in particular to the immunodiagnostics of allergic conditions.

The aim of the invention is to increase the sensitivity of the method.

The method is carried out as follows.

0.08 ml of blood is transferred with a sterile pipette into two Vidal tubes, one of which serves as a control, and the second with experience. In the test tube, 0.02 ml of the test allergen is placed in a test solution of 3% sodium citrate, and 0% in the control tube. , 02 mp of 5% citrate solution. Sodium (standard allergens are mixed with anticoagulant - 25% sodium citrate solution in a 4: 1 ratio, non-standard allergens are used in doses corresponding to the formulation, neutrophil damage test index. Then Pro, tags with a study image The blood cells are incubated in a thermostat at 45 - 46 ° C for two hours, after which smears are prepared and stained according to Mai-Grünwald, Papenheim or Romanovsky-Giemsa. The reaction counting is carried out by calculating the percentage of modified leukocytes in the experimental (N). and control (H) smears. The leukocyte sensitization index is determined by the formula

H i

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Evaluation of the results: negative - index О - 0.05; positive - index 0.06 0.35; sharply positive index more than 0.3.

Patient XD Patient X Diagnosis - chronic decompensated tonsillitis, frequent sore throats in the past history. The patient was admitted to the ENT clinic for tonsillectomy. On the day of admission, a determination of the sensitization of blood leukocytes to streptococcal (KNIIEM) and tissue allergens was carried out.

Blood obtained from the patient's finger by the standard method was transferred with a sterile pipette into three 0.08 ml vials, one of which served as a control, two others were experienced. 0.02 ml of 5% sodium citrate was placed in the control tube, well experienced

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investigated allergens (in the first — 0.02 i-ai serptococcal allergen on 5% sodium citrate solution, in the second — 0.02 ml of tissue allergen on sodium citrate solution). Then the test tubes with blood samples were incubated in a thermostat at 45 ° C for two hours, after which smears were prepared and stained according to May-Grunwald. When calculating, the percentage of cells with homogeneous basophilic cytoplasm in relation to leukocytes with invariable cytoplasm and cores was taken into account. In a test tube with a streptococcal allergen, the number of changed leukocytes was 31%, in a test tube with a tissue allergen - 47%, in a test tube 5%, Sensibi Index leukocyte size is determined by

H N

7Qo mule 1 is the percentage of altered leukocytes in the test smear; H - in the control, the Sensitization index to the streptococcal allergen was 0.26, tissue tissue - 0.42,

Thus, the result obtained is characteristic of patients with a depopensed form of chronic tone-, zillite. The performed pathological control confirmed the clinical diagnosis.

From the results of a comparative assessment of the effectiveness of the proposed and well-known (prototype) methods obtained in the study of 3 groups of patients, 20 people each, it follows that the proposed method for determining sensitization of leukocytes has significantly pronounced diagnostic advantages compared with the known method. in case of turculeuze, an increase in leukocyte damage index with turbeculin is 2.16 times (0.268 + 0.0835 ISC according to the proposed and 0.124 ± 0.0317 according to the prototype), in case of intolerance to streptomycin - 2.2 times (0.103f 0.0294 and 0.047 ± 0.0 122, respectively), with intolerance to novocaine, 1.35 times (0, .168 ± 0.0509 and 0.124 ± 0.0373, respectively).

The data presented show that the proposed method for determining the sensitization of leukocytes is universally suitable for the diagnosis of allergic processes, inductively coupled 5

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with infectious, nephrotic, and autogenic allergens, it is characterized by a high degree of specificity, a pronounced diagnostic sensitivity, and it can reveal allergic conditions not determined by known methods.

Thus, the potentials of the proposed method for determining sensitization of leukocytes include: increasing the diagnostic sensitivity of the proposed method in comparison with the prototype 1.2-2.16 times; simplification of the procedure for evaluating the results of the reaction and the treatment of blood smears, which makes available research in clinical laboratories; carrying out methods of setting and evaluating the results of the reaction in accordance with the pathogenetically leading factors of allergic

Editor T. Parfenova Order 6151/48

Compiled by V.Litovchenko Tehred L. Serdyukova

Circulation 776 VNIIPI USSR State Committee

for inventions and discoveries 113035, Moscow, Zh-35, Raushsk nab, 4/5

Production and printing company, Uzhgorod, Projecto st., 4

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inflammation proceeding under conditions of increased temperature reaction, allows to register all cells involved in the determination of the allergic status, regardless of the degree of their specificity

Claims (1)

Invention Formula The method of determining the sensitization of leukocytes, including the incubation of leukocytes in the blood of patients with an allergen, followed by the preparation of smears 5 blood and their staining, which differs with the fact that, in order to increase the sensitivity of the method, leukocytes with an allergen are incubated at 45-46 ° C, smears are stained 0 according to Mai-Grunwald and by the presence of cells with a homogeneous basophilic stain or their cytolysis, leukocyte sensitization is determined. Proofor VOUT ha Subscription