SU1120969A1 - Method of diagnosis of staphylococcal pneumonia of children at early age - Google Patents

Abstract

A METHOD FOR DIAGNOSTIC STAFILOCOCCAL PNEUMONIA IN CHILDREN OF EARLY AGE by examining blood, characterized in that, in order to improve the accuracy and shorten the time of diagnosis, lymphocytes derived from the patient's blood in the reaction of rosette, which sensitize with CPS-chytotectosetoctose, chibiococcalceptocytes, chytococcus, and chitosomas are diagnosed with rosiococcalceptocytes. : 20 for 25-30 minutes at 37-38 ° C and according to an increase in the number of rosetting lymphocytes, staphylococcal pneumonia is diagnosed.

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The invention relates to medicine, in particular to the diagnosis of staphylococcal pneumonia in young children. A known method for the diagnosis of staphylococcal pneumonia, such as bacteriological, involving the detection of staphylococci from biological materials (blood, sputum, etc.) followed by the determination of their toxigenicity 1. However, the widespread use of drugs (mainly antibiotics) led to a sharp decrease in the diagnostic value this way. A serological method of studying staphylococcal infection is known, consisting in the reaction of antigen-antibody in the presence of rabbit erythrocytes, according to the degree of hemolysis of which antibodies 2 are detected. However, this method is not sufficiently accurate due to the presence of primary and secondary immunodeficiency in young children time with its implementation. The purpose of the invention is to increase the accuracy and reduce the time of diagnosis. This goal is achieved in that according to the method of diagnosing staphylococcal pneumonia in young children, by examining blood, lymphocytes isolated from the patient's blood in a rosette reaction are examined, which sensitize with staphylococcal c-toxin at a dilution of 1: 15-1: 20 for 25-30 min at 37-38 ° C, and to increase the number of rosette lymphocytes, staphylococcal pneumonia is diagnosed. The method is carried out as follows. Lymphocyte sensitization. Take 3.0 ml of the patient's blood from the cubital vein into a tube with heparin diluted with saline 1:25 at the rate of 0.1 ml per 1 ml of blood. On the density gradient (1.077) of veragraphin-ficoll, lymphocytes are isolated from the obtained blood, they are washed three times with phosphate buffer (pH 7.2). Four samples of 0.1 ml (2 compared and 2 basic) were prepared from lymphocytes with the same lymphocyte concentration of 2 x 10 cells / ml. At the same time, a working suspension of sheep erythrocytes is prepared for determining T-cells. For this, the sheep erythrocytes are washed three times with phosphate buffer (pH-7.2) by centrifugation (at 1000 rpm), and 0.3% / red working cell suspension is prepared from the resulting dense precipitate. To determine B-lymphocytes, a working suspension of zymosanum is prepared by adding 1 ml of a guinea pig complement to a dilution of 1: 5 to 1.0 ml of 0.1 ° / o-zymosan. The resulting mixture was incubated at 37 ° C for 30 minutes. Then it is washed twice with a physiological solution of 5 min each by centrifugation at 1000 rpm. After adding 10 ml of phosphate buffer (pH 7.2) to the precipitate, a working suspension of zymosan is obtained to determine the lymphocytes. In two compared test tubes, the number of T and B lymphocytes is determined in a known manner. To two m of the main test tubes containing 0.1 ml each of lymphocyte suspension, 0.1 ml of staphylococcal ostoxin is added at a dilution of 1:15 -1: 20 depending on hemolytic activity. Short sensitization of lymphocytes is carried out by incubating the mixture in a thermostat at 37-38 ° C for 25-30 minutes. After incubation, both mixtures are washed three times with phosphate buffer (pH 7.2) and then the usual reaction is carried out to determine the number of T and B lymphocytes. The reaction is positive, since the number of T-lymphocytes increased by at least 10 rosette-forming cells when compared with the compared experience. Example I. Patient K., b months., Ist.bol. No. 1044, admitted with severe dyspnea (up to 50 times per minute), cyanosis of the nasolabial triangle at rest, cough of a wet nature, wheezing and crepitus in the lungs, enlarged liver size. Repeated blood cultures of staphylococcus growth were not given. On the first day, T- and B-lymphocytes were determined before and after incubation for 27 min with 1: 20-fold dilution with staphylococcal toxin and lymphocyte suspension. As a result, the number of rosette T-lymphocytes increased by 13 cells. The number of rosette B-lymphocytes remained unchanged. By the end of the second week of the disease, an increase in the level of alpha antitoxin in the patient's serum from 0.25 AU to 1.0 AE was detected. On the basis of the clinic and the increase in specificity of sensitized T-lymphocytes by 13 cells, the diagnosis was established on the first day: Staphylococcus pneumonia. The increase in the level of staphylococcal alpha antitoxin from 0.25 A.E. to 1.0 AE, as well as the effect of specific antistaphylococcal therapy, confirmed the validity of the diagnosis. Example 2. Sick D., 5 months, East. bol No. 2199, was admitted to the hospital with clinical manifestations of respiratory scarcity, a hyperthermic reaction to 39.2 °, intoxication, characterized by vesicular data (wheezing and crepitus) in the lungs.

On the first day of the patient's stay in the hospital, the number of rosette T and B lymphocytes was determined before and after incubating the lymphocyte suspension for 25 min with 1:13, 1:14 and 1: 15 times diluted with staphylococcal toxin. At the same time, the results of the analysis were not obtained in 1:13 and 1:14 dilutions due to full and partial hemolysis of red blood cells.

At a dilution of 1:15, the number of rosette T cells increased after incubation to 11 cells. The number of B-lymphocytes, both before the incubation and after it, remained at the same numbers.

On day 5, the results of blood culture were obtained on the day of admission, where a pathogenic hemolytic staphylococcus was detected.

By the end of the second week of the disease, an increase in the level of staphylococcal alpha antitoxin from 0.25 AU to 1.0 AE was revealed. On the basis of the clinic and the proposed diagnostic method on the first day, the patient was diagnosed: staphylococcus pneumonia, which later (on the 5th bacteriological, by the end of the second week of the disease, serological) confirmed the validity of the established diagnosis. The goal for dilution of staphylococcal toxin 1:13 and 1:14 is not achieved, but achieved when the reaction is performed in a diluted staphylococcal toxin 1:15.

Example 3. Patient 3 ,, 6 months, East. bol No. 1803, he was admitted to the hospital with clinical manifestations of respiratory failure, intoxication, hyperthermic reaction, and characteristic vesicular data from the lungs (wheezing and crepitus).

On the first day of admission to the hospital, the number of rosetting T and B lymphocytes was determined before and after 25 minutes of incubating the suspension of the patient's Lymphocytes from 1:20, 1:21, and 1: 22 times diluted with staphylococcal toxin.

At the same time, an increase in rosette T-lymphocytes occurred on 17 cells during incubation, suspension of lymphocytes with 1: 20 times diluted staphylococcal toxin. When incubating lymphocyte suspensions with 1:21 and 1: 22-fold diluted staphylococcal toxin, no increase in rosette-forming T-lymphocytes is detected. The number of B-lymphocytes remains unchanged after incubation at all of the indicated staphylococcal toxin dilutions.

On the 6th day of the patient's stay in the hospital, a positive result was obtained from the sowing conducted on the day of admission, where a pathogenic hemolytic staphylococcus was diagnosed.

By the end of the second week of the disease, an increase in the level of staphytes of bovine alpha antitoxin in serum from 0.25 ae to 1.0 AE was detected.

Based on the clinic and application

The proposed diagnostic method on the first day of illness was diagnosed with staphylococcal pneumonia. Afterwards, on the 6th day of staying in the hospital and by the end of the second week of the disease, the validity of the established diagnosis was bacteriologically and serologically confirmed. The goal was achieved with the use of diluted staphylococcal toxin 1:20.

Staphylococcal toxin dilution

5 1:21 and 1:22 or more does not give a positive effect of increasing the number of rosette-forming T-lymphocytes.

Example 4. Patient D., 2, 6 months, East. bol No. 287, was admitted for inpatient treatment in serious condition with severe

0 by staining, cyanosis of the nasolabial triangle, acrocyanosis, humorous nature, temperature reaction up to 39.4 °, participation of auxiliary muscles in the act of breathing, characteristic vesicular data (wheezing and crepitus) from the side of the lungs.

On the first day of admission to the hospital, the number of rosette-forming T-and B-lymphocytes was determined before and after incubating the suspension of lymphocytes with 1: 15-fold

0 diluted with staphylococcal toxin for 25 min. At the same time, an increase in the number of rosette T lymphocytes after incubation with staphylococcal toxin by 19 cells was revealed. No increase in B lymphocytes was detected.

5 On the 6th and 8th days of the patient's stay in the hospital, the results of the patient's blood seeding were obtained. At the same time, hemolytic staphylococcus of one phagovar was revealed. By the end of the second week of the disease,

0 increase in the level of staphylococcal alpha antitoxin in serum from 0.25 AU to 1.0 AE.

However, already during the first day of the patient’s stay in the hospital, a diagnosis was made on the basis of the Research Institute of the Clinic and the application of the proposed diagnostic method: Staphylococcus pneumonia. Subsequently, on days 6 and 8 and by the end of the second week of the disease, the validity of the established diagnosis was confirmed.

0 Example 5. Ill B., 1 g 4 months, East. bol No. 2134, arrived in grave condition with pronounced manifestations of respiratory failure, hyperthermic reaction, intoxication, a thicket of wet nature, characteristic changes in the lungs (wheezing and crepitus). On the first day the patient was admitted to the hospital, the number of rosette T and B lymphocytes was determined.

before and after incubation, lymphocyte suspension with 1: 15-fold diluted staphylococcal toxin for 25 minutes. At the same time, an increase in the number of rosette T-lymphocytes was detected during incubation with staphylococcal toxin by 17 cells. No increase in B lymphocytes was detected.

Repeated blood cultures of the patient for excretion of staphylococcus did not give a positive result.

By the end of the second week of the disease, an increase in the level of staphylococcal alpha-antitoxin from 0.25 AU to 1.0 AU was revealed in the patient's serum. However, already during the first day a diagnosis was made: staphylococcal pneumonia on the basis of the clinic and the application of the proposed diagnostic method.

The increase in the level of staphylococcal alpha antitoxin by the end of the second week of the disease and the effect of the specific antistaphylococcal therapy carried out confirmed the validity of the established diagnosis.

The possibility of revealing the staphylococcal etiology of pneumonia by the proposed method is shown under the condition of a negative bacteriological response.

Example 6. Patient 3. 2 months., East. bol Luo 1888, was admitted for inpatient treatment in severe condition with manifestations of respiratory failure, wet cough, hyperthermic reaction, tonic-clonic seizures, and characteristic changes (wheezing and crepitus) from the lungs.

On the first day of the patient's stay in the hospital, the number of rosette-forming T and B lymphocytes was determined before and after incubating the lymphocytes with 1: 20-fold diluted staphylococcal toxin for 25 minutes. At the same time, an increase in the number of T-lymphocytes after a short incubation with staphylococcal toxin by 11 cells was revealed. An increase in B lymphocytes was not detected.

On days 5 and 6 of the patient's inpatient stay, hemolytic staphylococci of one phagovar were inoculated from the blood. In the course of the disease, it was not possible to detect an increase in staphylococcal alpha antitoxin in the patient's serum.

However, already on the nerve day of the patient's stay in the hospital, on the basis of the clinic and the application of the proposed diagnostic method, the diagnosis was made: staphylococcus pneumonia.

The validity of the established diagnosis was confirmed on the 6th day of the patient's stay in the hospital by bacteriological method of research and effectiveness

antistaphylococcal therapy. The possibility of diagnosis was shown provided that a serological method of detecting the etiology of pneumonia is negative.

Example 7. Ill K., 5 months., East. / Bol. No. 1846, entered in serious condition for inpatient treatment. On admission, pronounced manifestations of respiratory failure, hyperthermic reaction, cough of wet nature, wheezing and crepitus in the lungs were noted.

On the first day the patient was admitted to the hospital, the number of rosette-forming T- and B-lymphocytes was determined before and after incubating the suspension of lim5 lymphocytes with 1: 20-fold diluted staphylococcal toxin for 25 minutes. At the same time, an increase in the number of rosette-forming T-lymphocytes was revealed after a short incubation with 21 cells of staphylococcal toxin. An increase in B-lymphocytes was not detected.

Repeated bacteriological examination of the patient's blood did not reveal the growth of staphylococci.

Determination of Staphylococcal Level

5 alpha-antitoxin also did not reveal its increase in the dynamics of the disease. On the first day of admission, the patient was diagnosed with staphylococcal disease on the basis of the clinic and the use of the proposed diagnostic method.

0 pneumonia. The effect of the specific antistaphylococcal therapy carried out confirmed the validity of the established diagnosis. The possibility of early diagnosis of staphylococcal etiology of pneumonia is shown under the condition of negative bacteriological and serological methods of diagnosis.

The table presents a comparative assessment of the accuracy of diagnosis of staphylococcal pneumonia in young children.

Q when the prototype method and the proposed method.

As can be seen from the presented data, an increase in the number of rosette T lymphocytes by more than 10 cells is significant for the diagnosis of staphylococci5 of the etiology of pneumonia in young children. At the same time, an increase in B-lymphocytes does not occur and is not informative. The diagnostic diagnostic test for this nosological unit.

Thus, the use of the proposed method for the diagnosis of staphylococcal pneumonia in young children provides, compared to known methods - bacteriological and serological reduction of the time of examination (10-12 days), increasing the accuracy of diagnosis (approximately 4 times).

Claims (1)

METHOD FOR DIAGNOSIS AND STAILOCOCCUS PNEUMONIA IN CHILDREN OF EARLY AGES by blood test, characterized in that, in order to increase accuracy and reduce the time of diagnosis, lymphocytes removed from the patient’s blood in the reaction of rosette formation are sensitized, which sensitize staphylococcal a-1 toxin 1 in dilution: : 20 for 25-30 minutes at 37-38 ° C and staphylococcal pneumonia is diagnosed by an increase in the number of rosette-forming lymphocytes.