Diagnosis of stress cardiomyopathy

FIELD: medicine, diagnosis, in particular earlier diagnosis of heart disorders in sportsmen.

SUBSTANCE: claimed method includes detecting of cardial antigen presence in blood serum using reaction of passive hemagglutination inhibition, and when titer of antimiocardial test-serum in presence of serum of tested subject by to ore more times stress cardiomyopathy is diagnosed.

EFFECT: diagnosis of stress cardiomyopathy in earlier steps.

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The proposed method relates to medicine, particularly cardiology, and can be used for early (pre-clinical) diagnosis of heart disease in athletes.

The most frequent disease of the heart in athletes is degeneration of the myocardium due to physical stress (1, 2). In the international classification of diseases (before revision) this disease is called stress cardiomyopathy, therefore, in recent years in our country instead of the term "degeneration of the myocardium due to physical stress" began to use the term "stress cardiomyopathy" (3, 4). The use of this term is justified also by the fact that patients is not only a physical and nervous stress.

The primary method of detection of stress cardiomyopathy is an ECG study. About the development of degenerative process in the myocardium show changes of repolarization and the emergence of rhythm and conduction. These changes are usually accompanied by a drop in performance. However, degeneration of the myocardium can be both diffuse and focal in nature, so in the latter case, changes on the ECG may be missing.

Therefore, the main disadvantage of ECG diagnosis is that it is not possible to identify early stages of development of stress CT is homeopatie before any changes the recorded ECG focal changes.

In connection with the foregoing, is warranted search different information criteria, indicating the development of myocardial degeneration, even when there are no ECG changes and does not suffer the performance, i.e. the pre-clinical stage of disease. Such early diagnosis of myocardial degeneration will allow to carry out treatment and preventive measures. This, in turn, gives the opportunity to maintain health, high performance and to prevent the development of disease.

The closest technical solution to the claimed method is a method described in the work Gavrilova E. "Stress cardiomyopathy in athletes: (dystrophy of the myocardium of physical stress)". Abstract. Prof. Diss., SPb., 2001.

According to this method, one of the criteria for the diagnosis of stress cardiomyopathy is the detection in serum of antibodies to the antigen of the myocardium in the reaction (TPHA) passive haemagglutination.

The objective of the invention is to increase the effectiveness of the treatment of stress cardiomyopathy by identifying it on the preclinical stages of development.

However, this method confirms the presence of myocardial degeneration by clinical stage, but you can't identify it at the preclinical stage, when netizenry on ECG, that's when you need to prevent that did not develop clinical stage: to reduce the volume and intesively training loads and to prescribe the drugs.

This object is achieved due to the fact that in the known method for the diagnosis of stress cardiomyopathy by serological analysis of blood serum, blood serum to determine the presence of cardiac antigen with response inhibition passive haemagglutination and at lower titers of therapy for anti-myocardial test serum in the presence of serum of the studied persons in two or more times diagnosed with stress cardiomyopathy.

The proposed method for the diagnosis of stress cardiomyopathy is based on serological analysis of blood, which detect the presence of myocardial antigen by the statement of response inhibition passive haemagglutination (RTPA).

When it runs in the blood serum of the subject (in the case of destructive processes in the myocardium) serologic method to detect the presence of antigen, the presence and the level of which indicates the degree of myocardial damage.

The method is based on the fact that the antigen of the myocardium, which may be present in the blood during stress cardiomyopathy, inhibits the interaction of the test antigen with antibodies in response inhibition passive haemagglutination. When it is determined the lower titers of diagnostic antimonial serum after incubation with the test serum. In these cases, the reduction of the titer of the antisera to one degree or more (for example, from 1:160 to 1:80 and over) mean detection in blood antigen and is a sign of destruction of the myocardium. The lower titers of test antisera on 2 stages and more allows you to assess the level of detectable antigen and, therefore, the degree of destruction of the myocardium.

The presence in the serum antigens myocardium indicates the presence of stress cardiomyopathy, even in cases when there are no clinical manifestations of the disease, or they are very poor, and when there are no ECG changes.

The proposed method preclinical diagnosis of stress cardiomyopathy is as follows.

Serological studies

1. Cooking myocardial antigen

The tissue of the myocardium from a person with 0 (I) blood group who died from accidental causes, used fresh or after storage in the frozen state.

A portion of the fabric is shredded with scissors, placed in a gauze bag, washed for one hour running water. The washed fabric is thoroughly crushed in a porcelain mortar with quartz sand, add saline, passed through one layer of cheesecloth to remove large particles. The resulting material is washed 5 times with saline by centrifugation at 18-20 thousand rpm for 1 minute. The precipitate is mixed with phosphate buffer solution (pH of 7.2) and incubated at 4°With during the day. After that, the liquid is centrifuged for 20 minutes at 3000 rpm./min, then adosados centrifuged at 18 tycobrahe 30 minutes at 4°C.

The supernatant was adjusted to a concentration of 2 mg protein in 1 ml and stored in the refrigerator with the addition of merthiolate in a dilution of 1:10000.

2. Getting therapy for anti-myocardial test serum with antibodies to myocardial antigen

Rabbits weighing not less than 2.5 kg are subjected to immunization with antigen from the myocardium according to the following scheme. In the pads of the hind legs of a rabbit injected with a mixture of antigen with complete adjuvant's adjuvant in a volume of 0.4 ml: 0.2 ml antigen containing 2 mg protein in 1 ml + 0.2 ml adjuvant. After 3 weeks antigen in a total dose of 10 mg injected directly into the enlarged lymph nodes. 10 days after the last immunization take the blood sample, determine serum antibody titer. When the titer of 1:320 and above blood samples were taken, prepare serum, which is heated at 37°C, poured into sterile vials and stored at 4°s to use.

3. Preparation of sera examined persons

The blood serum of patients with inactivating 30 minutes at 56°and drain the washed erythrocytes of sheep: 0.5 ml serum, 0.5 to sediment red blood cells, washed with physiological saline containing 1% krolic is her serum. Incubation for 2 hours at 37°and 18-20 hours at 4°C. the Supernatant is used for the given reaction.

4. Treponema pallidum haemagglutination assay. Preparation of red blood cells

Fresh, washed 3 times with saline, the sheep erythrocytes at a concentration of 50% is mixed with an equal volume of chromium chloride (3,8×10 M, 1 mg/ml), prepared ex tempore, with the addition of 100 μg/ml of protein.

After 5-minute incubation at room temperature the interaction is stopped by adding 20-50 volumes sabotinova saline solution and washed by centrifugation 4 times. After that erythrocytes resuspended in physiological solution containing 1% normal rabbit serum, receiving a 0.5% suspension of erythrocytes.

5. The setting reaction of the passive haemagglutination (TPHA)

Reaction set in tablets. Subjects serum of patients diluted by a factor of 2 in the wells containing 0.05 ml of 1% rabbit serum. Later in the wells add 0.025 ml of the suspension loaded with the antigen of red blood cells, incubated at 4°18-20 hours. Consider the reaction of agglutination of red blood cells in the negative control: 1% rabbit serum or 1% rabbit serum + normal human serum.

6. Formulation of response inhibition passive haemagglutination (RTPA)

0.05 ml of whole test blood serum of people who add in the hole, containing 0.05 ml of therapy for anti-myocardial test serum diluted 1:10. The mixture was incubated for 2 hours at a temperature of 37°and then 18-20 hours at 4°C. After that, the serum is diluted by a factor of 2, add to each well loaded with antigen, the red blood cells, determine the final dilution at which agglutination occurred, in comparison with dilutions therapy for anti-myocardial test serum serum, where the test material was added. The lower titers of therapy for anti-myocardial test serum by one step (for example, from 1:160 to 1:80) estimate as a likely indicator of the presence of the antigen of the damaged myocardium, decreased by 2-4 degrees as indicators of high antigen content of the myocardium, which indicates the presence of stress cardiomyopathy.

In control studies, the reduction of the titer of agglutination at 1 stage ensured the detection of 3 µg protein antigen, 2 stage - 6 µg, 4-speed - 12 µg protein antigen.

Clinical example.

Patient K. History no._____.

Professional athlete, master of sports of international class 18. Asked the sports medicine center complaining of reduced tolerance to physical activity, decreased athletic performance, no other complaints.

According to clinical examination and objective signs of disease were absent. Heart t is on rhythmic, correct, no noise, rhythm heart rate = 62 beats./min, BP = 120/70. In the lungs vesicular breathing, wheezing no. Peripheral edema no. ECG is normal. Pathological changes. Biochemical and clinical analysis of blood without pathological changes. An ultrasound of the heart pathology was not found.

The presence of cardiac antigen carried out by the statement of response inhibition passive haemagglutination. Results - lower titers of therapy for anti-myocardial test serum at stage 1 (from 1:160 to 1:80), which corresponds to the presence in the blood antigen damaged myocardium.

Diagnosed with stress cardiomyopathy

The patient is assigned therapy: Neoton 4 grams per day, Mildronate 10% - 10 ml/day, papansin 10 ml/day. therapy was carried out for seven days.

After the end of treatment the patient began intensive training. Tolerance to physical load fully recovered. Significantly increased athletic performance.

The example says that the patient had stress cardiomyopathy, pre-clinical stage. In this period did not reveal clinical manifestations and diagnosis made only by means of serological blood tests.

Studies suggest that the proposed method has) the public importance and can be used for the diagnosis of preclinical stage of stress cardiomyopathy. The proposed method can be used in medical practice for the diagnosis of stress cardiomyopathy its preclinical stage.

Thus, the proposed method allows to diagnose stress cardiomyopathy and to take urgent measures for future medical treatment that will prevent the development of disease and to maintain health and high performance. The advantage of this method is its sensitivity, high information content and specificity.

Sources of information

1. Butchenko L.A., Kushakovskij MS Dystrophy of the myocardium in athletes. - M.: Medicine. - 1980.

2. Dembo A.G. Medical control in sport. - M.: Medicine. - 1988.

3. Zemtsovsky EV Dystrophy of the myocardium in athletes // Cardiology. - 1994, No. 9. - P.65-74.

4. Zemtsovsky EV, Bondarev S.A., M. Lobanov Connective tissue dysplasia, tolerance to physical activity and degeneration of the myocardium physical overexertion // journal of sports medicine Russia. - 1999. - V.24, n 3. - P.27.

5. Gavrilova E.A. Stress cardiomyopathy in athletes: (dystrophy of the myocardium of overtraining), abstract. Prof. Diss., St. Petersburg,. 2001.

A method for the diagnosis of stress cardiomyopathy by serological analysis of blood serum, characterized in that the serum to determine the presence of cardiac antigen with response inhibition passive haemagglutination and at lower titers of therapy for anti-myocardial test serum in the presence of serum of the studied persons in two or more times diagnosed stress cardiomyopathy.



 

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