RU2557916C1 - Method of diagnosing occlusive affection of vessels in patients with cardiovascular diseases - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to the field of medicine and is intended for the diagnostics of an occlusive affection of vessels in patients with cardiovascular diseases. The age, the level of total homocysteine in blood, the presence of C677T polymorphism mutations in the gene of methyletetrahydrofolate reductase, factor V gene G1691A Leiden mutation, 675 4G/5G polymorphism mutations in the gene of the plasminogen activator inhibitor type I are determined in the patient, after which a value of the discriminant function is calculated by formula. In case of D>0, the presence of the occlusive affection of vessels is diagnosed, if D<0, the absence is diagnosed.

EFFECT: invention provides the effective method for the diagnostics of the occlusive affection of vessels in the patients with cardiovascular diseases.

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Description

The invention relates to medicine, namely to cardiovascular disease, and can be used to diagnose occlusive vascular lesions in patients with cardiovascular disease (CVD).

A known method for the diagnosis of vascular lesions in patients with CVD by biochemical examination of the level of total homocysteine (oGci) in blood plasma; with an oGcI content of more than 10 μmol / L, the possibility of vascular lesions is judged (Katrusiak AE, Paterson PG, Kamencic H., Shoker A., Lyin AW Pre-column derivatization high-performance liquid chromatographic method for determination of cysteine, cystenyl-glycine, homocysteine and glutathione in plasma and cell extracts // Journ. Chromatogr. B. Biomed. Sci. Appl. - 2001. - V.758, No. 2. - P.207-212).

The disadvantage of this method: the method does not allow to diagnose occlusive lesions in the vessels.

Studies are known aimed at creating additional laboratory tests to diagnose vascular lesions (thrombophilic disorders). These include studies of genetic polymorphisms of factor V - the Leiden mutation or the prothrombin G20210A mutation (Dahlback B. Inherited resistance to activated protein C, a major cause of venous thrombosis, is due to a mutation in the factor V gene // Haemostasis. - 1994. - Vol.24. - P.139), as well as other genetic markers of thrombophilia, such as MTHFR and 4G / 5G polymorphism of plasminogen activator inhibitor (Akar N., Akar E., Akcay R., Avcu F., Yalcin A. , Cin S. Effect of methylenetetrahydrofolate reductase 677 CT, 1298 AC, and 1317 TC on factor V 1691 mutation in Turkish deep vein thrombosis patients // Thromb Res. - 2000. - Vol. 97. - P.163-167).

The disadvantage of these studies is the lack of practical criteria that could be used as markers of occlusive vascular lesions in patients with CVD.

A known method for the diagnosis of occlusive vascular lesions in patients with CVD by duplex ultrasound examination - DUSI (National guidelines for the management of patients with vascular arterial pathology (Russian consensus document). Part 1. Peripheral arteries. - M .: Publishing House NTSSSH im. A. N. Bakuleva RAMS, 2010 .-- 176 p.). This method is accepted by the authors as a prototype.

The disadvantage of the prototype is that each study is aimed at diagnosing a specific vessel, while there is no criterion that allows you to diagnose the state of the vessels of the body as a whole.

The technical result of the invention is the expansion of the arsenal of tools for the diagnosis of occlusive vascular lesions in patients with CVD, the possibility of detecting occlusive vascular conditions of the body as a whole.

The specified technical result is achieved by the fact that in the method for diagnosing occlusive vascular lesions in patients with cardiovascular diseases, according to the invention, the level of total homocysteine in the blood, the presence of mutations of the C677T polymorphism in the methylene tetrahydrofolate reductase gene, the Leiden mutation G1691A in the factor V gene, mutations of the 675 4G polymorphism are determined / 5G in the plasminogen activator inhibitor gene of type I and the age of the patient is taken into account, then the value of the discriminant function is calculated by the formula:

D = -0.531 × B-15.466 × MTHFR + 19.0338 · FV Leiden + 0.9814 · (PAI-1) + 0.89 · oGci-18.266,

Where:

where B is the patient's age, years;

MTHFR - mutation of C677T polymorphism in the methylene tetrahydrofolate reductase gene: 0 - normal, 1 - heterozygous for mutation, 2 - homozygous for mutation;

FV Leiden - Leiden mutation G1691A in the factor V gene: 0 - normal, 1 - heterozygous for the mutation, 2 - homositis for the mutation.

PAI-1 - mutation of polymorphism 675 4G / 5G in the gene of plasminogen activator inhibitor type I: 0 - normal, 1 - heterozygous for mutation, 2 - homozygous for mutation;

oGci - the level of total homocysteine, µmol / l;

and with a value of D> 0, the presence of occlusive vascular lesions is diagnosed, and with D <0, the absence.

The proposed method is based on an analysis of the results of a study of 46 patients with CVD suffering from vascular damage of the whole organism of varying severity. For the study, two groups of patients were formed. The first group (21 people) consisted of patients with occlusive lesions of blood vessels of the body of various localization (pulmonary arteries, coronary arteries, deep veins of the lower extremities, brachiacephalic arteries, retinal vessels, sprays of the abdominal cavity). The second group (25 people) consisted of patients without occlusive vascular lesions with hemodynamically insignificant vascular lesions in the form of endothelial disorders and / or hemodynamically insignificant stenosis. The age of the patients of the first group was 45.7 ± 14.03 years, the age of the patients of the second group was 59.5 ± 8.9 years. By gender distribution, men predominated in both groups. In the first group - 15 men and 6 women; in the second group - 13 men and 12 women. Diagnoses of diseases of all patients were verified clinically and using neuroimaging methods: magnetic resonance imaging and / or spiral computed tomography of the brain in angiography. In addition, ultrasound diagnostics of the brachiocephalic arteries (USDA BCA), transcranial dopplerography (TCD), duplex ultrasound (DUS) of the vessels of the lower extremities, cerebral vessels and neck were performed. The study of neurological status was carried out according to the methodology of neurological examination (Skoromets A.A., Skoromets T.A. Topical diagnosis of diseases of the nervous system: a Guide for doctors. - 3rd ed., Rev. And add. - St. Petersburg: Polytechnic, 2000 . - 399 p .: silt).

To identify the most significant correlations in the pathology of vascular lesions, we studied 25 indicators, including: age, the presence of concomitant chronic diseases of the cardiovascular, respiratory, endocrine systems, the presence of neurological syndromes, oGci, polymorphism of the L 33 P (T> C) platelet receptor gene (ITGB3) (glycoprotein 3a), 807 C> T polymorphism (F224F) of the platelet receptor gene (ITGA2), C677T polymorphism in the methylenetetrahydrofolate reductase gene (MTHFR), mutation G1691A (Arg506Gln) in the gene for factor V (FV Lemen polymorphism F75) 4G / 5G in the gene inhibits pa I-type plasminogen activator (PAI-1), 455 Polymorphism G / A fibrinogen gene (FCB).

The following variables turned out to be the most significant as a result of discriminant analysis: patient age, mutation of C677T polymorphism in methylene tetrahydrofolate reductase gene, Leiden mutation G1691A in factor V gene, 675 4G / 5G polymorphism mutation in type I plasminogen activator inhibitor gene (PAI-1) and level total homocysteine.

The percentage of correct classification (diagnostic accuracy) was 88.4%; for the group with occlusive vascular lesions - 96.7%; for the group with hemodynamically insignificant stenosis - 78.7%.

The method is carried out, for example, as follows.

After conducting a clinical trial and a neurological examination (Skoromets A.A., Skoromets T.A. Topical diagnosis of diseases of the nervous system: a Guide for doctors. - 3rd ed., Rev. And add. - St. Petersburg: Polytechnic, 2000. - 399 s.: Ill) a patient with CVD is referred for laboratory testing. A study of GcI in blood plasma is performed by means of an HPLC analysis on an Agilent 1100 chromatograph with photometric detection (Laboratory diagnostic technology for detecting hyperhomocysteinemia. ”Zhloba AA, Blashko EL, Nikitina VV, No. FS 2009/309 from 09/04/09). A genetic study is being carried out to detect mutations of C677T polymorphism in the methylene tetrahydrofolate reductase gene, Leiden mutations G1691A in the factor V gene, and 675 4G / 5G polymorphism mutations in the type I plasminogen activator inhibitor gene. Then, based on the results of the studies, taking into account the age of the patient, the value of the discriminant function is calculated according to the formula:

D = -0.531 × B-15.466 × MTHFR + 19.0338 · FV Leiden + 0.9814 · (PAI-1) + 0.89 · oGci-18.266

and with a value of D> 0, the presence of occlusive vascular lesions is diagnosed, and with D <0, the absence.

The method is illustrated by the following clinical examples.

Example 1. Patient B., 64 g. Diagnosis: encephalopathy of mixed origin (discirculatory, dysmetabolic). Initial atherosclerotic disorders of the cerebral arteries. Varicose veins of the lower extremities. Hyperhomocysteinemia of moderate severity. A study of the proposed method. Biochemical study of blood plasma: the level of oGci - 14.2 μmol / L.

Molecular genetic study:

the presence of C677T polymorphism mutations in the methylene tetrahydrofolate reductase gene is the norm (MTHFR = 0),

the presence of Leiden mutations G1691A in the factor V gene is normal (FV Leiden = 0),

the presence of mutations in 675 4G / 5G polymorphism in the gene of type I plasminogen activator inhibitor is normal (PAI-1 = 0).

The value of the discriminant function according to the formula, taking into account age:

D = -0.531 × B-15.466 × MTHFR + 19.0338 · FV Leiden + 0.9814 · (PAI-1) + 0.89 · oGci-18.266

D = -39.612; D <0 - diagnose the absence of occlusive vascular lesions, which was confirmed by ultrasound of vessels of the lower extremities, the main arteries of the body.

Example 2. Patient M., 46 years old. Diagnosis: Varicose veins of the lower extremities. Residual effects after thromboembolism of small branches of the pulmonary artery. Heart failure II f.k. by NYHA. Hypertension III art, hypertension 3 art, the risk of MTR 3. Dyscirculatory encephalopathy I art. Hyperhomocysteinemia of moderate severity.

A study of the proposed method. Biochemical study of blood plasma: the level of oGci - 18.5 μmol / L.

Molecular genetic study:

the presence of C677T polymorphism mutations in the methylene tetrahydrofolate reductase - heterozygote gene (MTHFR = 0)

Leiden mutations G1691A in factor V gene - homozygote detected (FV Leiden = 2)

Mutations of 675 4G / 5G polymorphism in the gene of type I plasminogen activator inhibitor were not detected (PAI-1 = 0).

The value of the discriminant function according to the formula, taking into account age:

D = -0.531 × B-15.466 × MTHFR + 19.0338 · FV Leiden + 0.9814 · (PAI-1) + 0.89 · oGci-18.266

D = 11.84; D> 0 - diagnose the presence of occlusive vascular lesions of the body. The diagnosis of the disease was verified using a clinical study, the results of ultrasound diagnostics of the vessels of the lower extremities, computed tomography of the chest. Ultrasound of vessels of the lower extremities: valve failure of the deep veins of the lower extremities. Computed tomography of the chest: thromboembolism of small branches of the pulmonary artery from two sides.

Example 3. Patient, 41 g. Diagnosis: Hypertension II Art., AH 2 Art., The risk of MTR 3. Coronary heart disease, angina pectoris 2 f. class. Dyscirculatory encephalopathy II Art. Hyperhomocysteinemia of moderate severity. A study of the proposed method. Biochemical study of blood plasma: the level of oGci - 15.3 mmol / L.

Molecular genetic study:

Leiden mutations G1691A in the gene of factor V - homozygous was found (FV Leiden - 2).

Mutations of 675 4G / 5G polymorphism in the gene of type I plasminogen activator inhibitor were not detected (PAI-1 = 0).

the presence of mutations in the C677T polymorphism in the methylenetetrahydrofolate reductase – heterozygous gene (MTHFR = 0).

The value of the discriminant function according to the formula, taking into account age:

D = -0.531 × B-15.466 × MTHFR + 19.0338 × FVLeiden + 0.9814 × (PAI-1) + 0.89 ° C-18.266 D = 11.65; D> 0 - the patient is diagnosed with occlusion of the vessels of the patient, which was confirmed by the performed ultrasonography of BCA, TCD. Found: hemodynamically significant stenosis of the arteries of the brain (70%), subcritical stenosis.

The proposed method expands the arsenal of tools for the diagnosis of occlusive vascular lesions in patients with CVD, makes it possible to diagnose the occlusive vessels of the body as a whole.

Claims (1)

A method for the diagnosis of occlusive vascular lesions in patients with cardiovascular diseases, characterized in that they determine the level of total homocysteine in the blood, the presence of mutations of the C677T polymorphism in the gene of methylenetetrahydrofolate reductase, the Leiden mutation G1691A in the gene of factor V, mutations of polymorphism 675 4G / 5G in the inhibitory gene plasminogen activator type I and take into account the age of the patient, then calculate the value of the discriminant function according to the formula:
D = -0.531 × B - 15.466 × MTHFR + 19.0338 × FV Leiden + 0.9814 × (PAI-1) + 0.89 × GHz - 18.266,
Where:
where B is the patient's age, years;
MTHFR - mutation of C677T polymorphism in the methylene tetrahydrofolate reductase gene: 0 - normal, 1 - heterozygous for mutation, 2 - homozygous for mutation;
FV Leiden - Leiden mutation G1691A in the factor V gene: 0 - normal, 1 - heterozygous for mutation, 2 - homozygous for mutation;
PAI-1 - mutation of polymorphism 675 4G / 5G in the gene of plasminogen activator inhibitor type I: 0 - normal, 1 - heterozygous for mutation, 2 - homozygous for mutation;
oGci - the level of total homocysteine, µmol / l;
and with a value of D> 0, the presence of occlusive vascular lesions is diagnosed, and with D <0, the absence.