Polypeptide marker for arteriosclerosis diagnosis

FIELD: medicine.

SUBSTANCE: there is offered an arteriosclerosis diagnostic technique based on the detection of various combinations of specific polypeptide markers. To detect the presence or absence of polypeptide marker, capillary electrophoresis, gaseous-phase ion spectrometry and/or mass spectrometry are used.

EFFECT: method allows early high-probability minimally invasive arteriosclerosis detection.

13 cl, 1 ex, 5 tbl

 

The present invention relates to the use factor of the presence or absence of one or more peptide markers in a sample of an individual for the diagnosis of arteriosclerosis, as well as to a method of diagnosis of arteriosclerosis, and the presence or absence of the peptide marker or peptide markers is an indication of the presence of arteriosclerosis.

Arteriosclerosis or atherosclerosis, calcification of blood vessels, respectively artery is the most frequent painful change vessels. Arteriosclerosis is a change in the blood vessels that occurs over many years and first flows unrecognizable. On the walls of blood vessels accumulate contained in the blood fatty substances and leukocytes (belascoaran), vessels are calcined, lose their elasticity and increasingly narrows the diameter of the vessels. As risk factors for the occurrence of arteriosclerosis believe hypercholesterolemia, high blood pressure, Smoking, diabetes mellitus, obesity, low physical activity and low social status, which, however, closely linked to other factors. Discuss other risk factors, such as hypertriglyceridemia, elevated blood concentrations of lipoprotein (a), homocysteine and certain parameters of inflammation, as with the reaction the active protein (CRP) or fibrinogen, and pyloric infection Chlamidien or Helicobacter.

Arteriosclerosis a long time do not cause any symptoms. Only when the diameter of the vessel at the expense of plaques clearly decreases or in the field of plaques formed a blood clot (thrombus), there are symptoms. The most frequent arteriosclerotic cardiac and vascular diseases are coronary heart disease, cerebrovascular disease and peripheral arterial occlusive disease, life-threatening complication of which, such as myocardial infarction, stroke or loss of the lower extremities, in Germany there are very often compared to poorer countries and are the cause of extremely high health care costs.

Due to arteriosclerosis, cardiovascular disease, also called cardiovascular disease, are in first place in the statistics of causes of death in Germany. So, cardiovascular disease in 1999 according to the Federal Statistical Office of Germany amounted to 48% of the most frequent causes of death, and ischemic (coronary) heart disease accounted for 21% and diseases of the cerebrovascular system was 10%. Compared to this, the proportion of cancer fatalities amounted to 26%, in the case of diseases of the respiratory system and the digestive system is s share was 6%, respectively 5%.

Although arteriosclerotic changes often occur in adolescence, clinical manifestations usually occur in middle and older age.

Arteriosclerosis cannot be treated with drugs, but only you can avoid it by prevention. Already begun vascular calcification cannot be destroyed, rigid vessel walls cannot be returned elasticity. However, you can clearly slow the progression of arteriosclerosis that influence the risk factors through lifestyle changes, for example by nicotine abstinence or drug (for example, using acetylsalicylic acid). Along with this, in the case of severe arterial vascular calcification there are also surgical methods, such as balloon dilatation (MOUTH, percutaneous intraluminal angioplasty, bypass surgery and stent use.

As previously reported, arteriosclerotic change cannot be treated therapeutically, and only you can prevent or slow down, especially important is the early recognition arteriosclerotic changes.

Under normal conditions only the diagnosis of the disease, which is a consequence of the previous one, allows the conclusion that the affected individual is sick arteriosclerosis. At the present time for this reveal narrowing of the blood vessels in the example, using a contrast medium or x-rays or by means of ultrasound. Moreover, usually examine the risk factors, like high cholesterol levels or diabetes. All of these methods, however, are insufficient for early and accurate diagnosis of arteriosclerosis. In particular there is a need to, if possible, minimally invasive, quick and requires less early detection of arteriosclerosis.

Now unexpectedly found that certain peptide markers in a sample of an individual can be used for diagnosis of arteriosclerosis.

Therefore, an object of the present invention is the use factor of the presence or absence of at least one polypeptide marker in a sample of an individual for the diagnosis of arteriosclerosis, and the polypeptide marker selected from the polypeptide markers№ 1, № 2, № 3, № 4, № 5, № 6, № 7, № 8 or No. 9, which are listed in table 1 values of molecular weight:

Table 1
Polypeptide markers for the diagnosis of arteriosclerosis and their molecular masses
Polypeptide marker No.Molecular weight [Yes]
2511,8
21864,7
32799,8
41340,6
51422,8
65882,0
71397,4
81886,6
94642,6

When using the present invention can be very early to diagnose arteriosclerosis. Due to this disease can be treated at an early stage or before its manifestations with well-known drugs and, thus, weaken its further course. According to the invention, further, it becomes possible which requires less cost, quick and reliable diagnosis of arteriosclerosis in part without or only minimal invasive procedures.

According to a special variant of the invention, the polypeptide marker is characterized not only by its molecular weight, but also its migration time (see table 2):

Table 2
Polypeptide markers for the diagnosis of arteriosclerosis, and the molecular masses and migration times
Polypeptide marker No.Molecular weight [Yes]The migration time [min]
12511,826,4
21864,749,2
32799,846,0
41340,666,7
51422,861,2
65882,033,3
71397,424,9
81886,644,7
94642,637,5

The migration time determined by capillary electrophoresis (capillary electrophoresis, CE), as indicated, for example, in the example in paragraph 2. Using a glass capillary with a length of 90 cm with internal d is amerom (ID) of 75 μm and an outer diameter (OD) of 360 μm at a voltage of 30 kV. As the solvent for the sample using 30% methanol and 0.5% formic acid in water.

It is known that migration in capillary electrophoresis can vary. However, the consistency with which suiryudan polypeptide markers, if any used CE systems are usually the same. To balance out the differences in time migration, the system can normalize using standards with known times of migration. These standards can be, for example, specified in the examples, the polypeptides (see example, paragraph 3, "Standards for CE-measurement").

Characterization of polypeptide markers listed in tables 1-3, are determined by capillary electrophoresis - mass spectrometry (capillary electrophoresis - mass spectrometry, CE-MS)method, which is described, for example, Neuroff and others (Rapid Communications in mass spectrometry, 20, 149-156 (2004)). The variation of molecular masses between individual measurements or between different mass spectrometers is relatively small, it usually is in the range of ±0.1%, preferably within ±0.05%.

Proposed according to the invention the polypeptide markers are proteins or peptides or cleavage products of proteins or peptides. They can be chemically modified, for example, by post-translational modifications like glycosylation, phosphoryl is the formation, alkylation or the formation of disulfide bonds, or can be modified by other reactions, for example, in the framework of the cleavage reaction. In addition, the polypeptide markers can also be chemically modified in the framework cleanup of samples, for example oxidized.

On the basis of parameters that define polypeptide markers (molecular weight and migration), by known prior art methods it is possible to identify the sequence of the corresponding polypeptide.

Proposed according to the invention polypeptides (see table 1 or 2) is used for diagnosis of arteriosclerosis. Under the diagnosis understand the recognition process by identifying symptoms or phenomena of the disease or lesion. In this case, the presence or absence of particular polypeptide markers make a conclusion about the presence of arteriosclerosis. For this sample of an individual identify proposed according to the invention the polypeptide markers, and their presence or absence can make a conclusion about the presence of arteriosclerosis. The presence or absence of a polypeptide marker can be determined by any known prior art method. Methods that can be used to specify, for example, next.

Polypeptide marker is present when the measured value is at least the mayor as high as the threshold value. If the measured value is below the threshold value, the polypeptide marker is absent. The threshold value can be defined either by the high sensitivity of the measuring method (limit of detection), or you can find by experiment.

According to the present invention, the threshold value preferably is exceeded, when the measured value of the sample in respect of a particular molecular weight is at least twice higher than that blank sample (for example, only the buffer or solvent), more preferably a measured value at least three times higher, even more preferably at least four times higher in the highest degree preferably five times higher than that blank sample.

Polypeptide, the polypeptide marker or markers are used in such a way that determine its/their presence or absence and the presence or absence is indicative of arteriosclerosis. So, there are polypeptide markers which are usually present in patients with arteriosclerosis (the patient), as, for example, the polypeptide markers № 1, № 7, № 8, № 9, but not the basis of reliable paternal without arteriosclerosis (control). Moreover, there are polypeptide markers which are available to individuals without atherosclerosis (control), but more rarely or configuration but does not occur in individuals with arteriosclerosis. This is, for example, the polypeptide markers№ 2, № 3, № 4, № 5 and No. 6.

Table 3
Polypeptide markers for the diagnosis of arteriosclerosis, their molecular masses and migration times, and their presence or absence in patients with arteriosclerosis patients and controls
(preparation of samples and the definition as described in the example)
Polypeptide marker No.Molecular weight [Yes]The migration time [min]sickcontrolΔ
12511,826,40,600,000,60
21864,749,20,200,780,58
32799,846,00,200,780,58
41340,6 66,70,100,670,57
51422,861,20,000,560,56
65882,033,30,000,560,56
71397,424,90,500,000,50
81886,644,70,500,000,50
94642,637,50,500,000,50
sickthe proportion of individuals with arteriosclerosis, in which the polypeptide marker present in the sample
controlthe proportion of individuals without atherosclerosis, which polypeptide the th marker present in the sample
Δin the amount of (patient-control)

The individual from whom the sample, which determines the presence or absence of one or more polypeptide markers can be any individual who may suffer from arteriosclerosis, for example an animal or a person. Preferably in the case of the individual we are talking about a mammal, such as, for example, dog, horse, cat, and highly preferably people.

According to a preferred variant embodiment of the invention for diagnosing arteriosclerosis use not only one polypeptide marker, but a combination of polypeptide markers, and their presence or absence can be done a posteriori conclusion about the presence of arteriosclerosis. By comparing a variety of polypeptide markers can reduce or avoid falsification of the final result at the expense of the individual deviations from the typical probability of the presence of the patient or control of the individual.

According to a preferred variant of the invention, therefore, use two polypeptide marker is listed in table 1 or 2 for the diagnosis of arteriosclerosis. In particular, this combination of polypeptide markers:

- № 1 and № 2, № 1 and № 3, № 1 and the 4, № 1 and № 5, № 1 and № 6, № 1 and № 7, № 1 and № 8, № 1 and № 9;

- № 2 and № 3, № 2 and № 4, № 2 and № 5, № 2 and № 6, № 2 and № 7, № 2 and № 8, № 2 and № 9;

- № 3 and № 4, № 3 and № 5, № 3, № 6, № 3 and № 7, № 3 and № 8, № 3 and № 9;

- № 4 and № 5, № 4 and № 6, № 4 and № 7, № 4 and № 8, № 4 and № 9;

- № 5 and № 6, № 5 and № 7, № 5 and № 8, № 5 and № 9;

- #6 and # 7, # 6 and # 8, # 6 and # 9;

No. 7 and No. 8, No. 7 and No. 9; or

No. 8 and No. 9.

According to another preferred variant of the invention, the use of three polypeptide marker is listed in table 1 or table 2 for the diagnosis of arteriosclerosis. In particular, this combination of polypeptide markers:

No. 1 and No. 2 in combination with one of the polypeptide markers№ 3, № 4, № 5, № 6, № 7, № 8 or # 9;

No. 1 and No. 3 in combination with one of the polypeptide markers№ 4, № 5, № 6, № 7, № 8 or # 9;

No. 1 and No. 4 in combination with one of the polypeptide markers№ 5, № 6, № 7, № 8 or # 9;

No. 1 and No. 5 in combination with one of the polypeptide markers No. 6, No. 7, No. 8 or No. 9;

No. 1 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 1 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 1 and No. 8 in combination with a polypeptide marker No. 9;

No. 2 and No. 3 in combination with one of the polypeptide markers№ 4, № 5, № 6, № 7, № 8 or # 9;

No. 2 and No. 4 in combination with one of the polypeptide markers№ 5, № 6, № 7, № 8 or # 9;

No. 2 and No. 5 in kombinats and with one of the polypeptide markers No. 6, No. 7, No. 8 or No. 9;

No. 2 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 2 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 2 and No. 8 in combination with # 9;

No. 3 and No. 4 in combination with one of the polypeptide markers№ 5, № 6, № 7, № 8 or # 9;

No. 3 and No. 5 in combination with one of the polypeptide markers No. 6, No. 7, No. 8 or No. 9;

No. 3 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 3 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 3 and No. 8 in combination with a polypeptide marker No. 9;

No. 4 and No. 5 in combination with one of the polypeptide markers No. 6, No. 7, No. 8 or No. 9;

No. 4 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 4 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 4 and No. 8 in combination with a polypeptide marker No. 9;

No. 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 5 and No. 8 in combination with a polypeptide marker No. 9;

No. 6 and No. 7 in combination with a No. 8 or No. 9;

No. 6 and No. 8 in combination with No. 9; or

No. 7 and No. 8 in combination with No. 9.

According to another preferred variant of the invention, the use of four polypeptide marker is listed in table 1 or table is itzá 2 for diagnosis of arteriosclerosis. In particular, this combination of polypeptide markers:

- No. 1, No. 2 and No. 3 in combination with one of the polypeptide markers№ 4, № 5, № 6, № 7, № 8 or # 9;

- No. 1, No. 2 and No. 4 in combination with one of the polypeptide markers№ 5, № 6, № 7, № 8 or # 9;

- No. 1, No. 2 and No. 5 in combination with one of the polypeptide markers No. 6, No. 7, No. 8 or No. 9;

- No. 1, No. 2 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

- No. 1, No. 2 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- No. 1, No. 2 and No. 8 in combination with # 9;

- No. 1, No. 3 and No. 4 in combination with one of the polypeptide markers№ 5, № 6, № 7, № 8 or # 9;

- No. 1, No. 3 and No. 5 in combination with one of the polypeptide markers No. 6, No. 7, No. 8 or No. 9;

- No. 1, No. 3 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

- No. 1, No. 3 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- No. 1, No. 3 and No. 8 in combination with # 9;

- No. 1, No. 4 and No. 5 in combination with one of the polypeptide markers No. 6, No. 7, No. 8 or No. 9;

- No. 1, No. 4 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

- No. 1, No. 4 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- No. 1, No. 4 and No. 8 in combination with # 9;

- No. 1, No. 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

- No. 1, No. 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 1, who is 5, and # 8 in combination with No. 9;

- No. 1, No. 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- No. 1, No. 6 and No. 8 in combination with # 9;

- No. 1, No. 7 and No. 8 in combination with # 9;

No. 2, No. 3 and No. 4 in combination with one of the polypeptide markers№ 5, № 6, № 7, № 8 or # 9;

No. 2, No. 3 and No. 5 in combination with one of the polypeptide markers No. 6, No. 7, No. 8 or No. 9;

No. 2, No. 3 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 2, No. 3 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 2, No. 3 and No. 8 in combination with # 9;

No. 2, No. 4 and No. 5 in combination with one of the polypeptide markers No. 6, No. 7, No. 8 or No. 9;

No. 2, No. 4 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 2, No. 4 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 2, No. 4 and No. 8 in combination with # 9;

No. 2, No. 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 2, No. 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 2, No. 5 and No. 8 in combination with # 9;

No. 2, No. 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 2, No. 6 and No. 8 in combination with # 9;

No. 2, No. 7 and No. 8 in combination with # 9;

No. 3, No. 4 and No. 5 in combination with one of the polypeptide markers No. 6, No. 7, No. 8 or No. 9;

No. 3, No. 4 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 3, No. 4 and No. 7 in to the binachi with one of the polypeptide markers No. 8 or No. 9;

No. 3, No. 4 and No. 8 in combination with # 9;

No. 3, No. 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 3, No. 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 3, No. 5 and No. 8 in combination with # 9;

No. 3, No. 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 3, No. 6 and No. 8 in combination with # 9;

No. 3, No. 7 and No. 8 in combination with # 9;

No. 4, No. 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 4, No. 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 4, No. 5 and No. 8 in combination with # 9;

No. 4, No. 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 4, No. 6 and No. 8 in combination with # 9;

No. 4, No. 7 and No. 8 in combination with # 9;

No. 5, No. 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 5, No. 6 and No. 8 in combination with # 9;

No. 5, No. 7 and No. 8 in combination with No. 9; or

No. 6, No. 7 and No. 8 in combination with No. 9.

According to another preferred variant of the invention, the use of five polypeptide markers listed in table 1 or table 2 for the diagnosis of arteriosclerosis. In particular, this combination of polypeptide markers:

- № 1, № 2, № 3 and № 4 in combination with one of the polypeptide markers№ 5, № 6, № 7, № 8 or # 9;

- № 1, № 2, № 3 and № 5 in combination with one of the polypeptide markers No. 6 No. 7, No. 8 or No. 9;

- No. 1, No. 2, No. 3 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

- № 1, № 2, № 3 and № 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- No. 1, No. 2, No. 3 and No. 8 in combination with a polypeptide marker No. 9;

- No. 1, No. 2, No. 4 and No. 5 in combination with one of the polypeptide markers No. 6, No. 7, No. 8 or No. 9;

- No. 1, No. 2, No. 4 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

- No. 1, No. 2, No. 4 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- No. 1, No. 2, No. 4 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 5 and № 6 in combination with one of the polypeptide markers No. 7, No. 8 or No. 9;

- No. 1, No. 2, No. 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- No. 1, No. 2 and No. 5, No. 8 in combination with a polypeptide marker No. 9;

- No. 1, No. 2 and No. 6, No. 7 in combination with one of the polypeptide markers No. 8 or 3 9;

- No. 1, No. 2, No. 6 and No. 8 in combination with a polypeptide marker No. 9;

- No. 1, No. 2, No. 7 and No. 8 in combination with a polypeptide marker No. 9;

- No. 1, No. 3, No. 4 and No. 5 in combination with one of the polypeptide markers No. 6, No. 7, No. 8 or No. 9;

- No. 1, No. 3, No. 4 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

- No. 1, No. 3, No. 4 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- No. 1, No. 3, No. 4 and No. 8 in combination with a polypeptide marker No. 9;

No. 1, the 3, No. 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

- No. 1, No. 3, No. 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- No. 1, No. 3, No. 5 and No. 8 in combination with a polypeptide marker No. 9;

- No. 1, No. 3, No. 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- No. 1, No. 3, No. 6 and No. 8 in combination with a polypeptide marker No. 9;

- No. 1, No. 3, No. 7 and No. 8 in combination with a polypeptide marker No. 9;

- No. 1, No. 4, No. 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

- No. 1, No. 4, No. 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- No. 1, No. 4, No. 5 and No. 8 in combination with a polypeptide marker No. 9;

- No. 1, No. 4, No. 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- No. 1, No. 4, No. 6 and No. 8 in combination with a polypeptide marker No. 9;

- No. 1, No. 4, No. 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 5, № 6 and № 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 5, № 6 and № 8 in combination with a polypeptide marker No. 9;

- No. 1, No. 5, No. 7 and No. 8 in combination with a polypeptide marker No. 9;

- No. 1, No. 6, No. 7 and No. 8 in combination with a polypeptide marker No. 9;

No. 2, No. 3, No. 4 and No. 5 in combination with one of the polypeptide markers No. 6, No. 7, No. 8 or No. 9;

No. 2, No. 3, No. 4 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 2 No. 3 No. 4 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 2, No. 3, No. 4 and No. 8 in combination with a polypeptide marker No. 9;

No. 2, No. 3, No. 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 2, No. 3, No. 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 2, No. 3, No. 5 and No. 8 in combination with a polypeptide marker No. 9;

No. 2, No. 3, No. 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 2, No. 3, No. 6 and No. 8 in combination with a polypeptide marker No. 9;

No. 2, No. 3, No. 7 and No. 8 in combination with a polypeptide marker No. 9;

No. 2, No. 4, No. 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 2, No. 4, No. 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 2, No. 4, No. 5 and No. 8 in combination with a polypeptide marker No. 9;

No. 2, No. 4, No. 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 2, No. 4, No. 6 and No. 8 in combination with a polypeptide marker No. 9;

No. 2, No. 4, No. 7 and No. 8 in combination with a polypeptide marker No. 9;

No. 2, No. 5, No. 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 2, No. 5, No. 6 and No. 8 in combination with a polypeptide marker No. 9;

No. 2, No. 5, No. 7 and No. 8 in combination with a polypeptide marker No. 9;

No. 2, No. 6, No. 7 and No. 8 in combination with a polypeptide marker No. 9;

No. 3, No. 4, No. 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 3, No. 4, No. 5 and No. 7 the combination with one of the polypeptide markers No. 8 or No. 9;

No. 3, No. 4, No. 5 and No. 8 in combination with a polypeptide marker No. 9;

No. 3, No. 4, No. 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 3, No. 4, No. 6 and No. 8 in combination with a polypeptide marker No. 9;

No. 3, No. 4, No. 7 and No. 8 in combination with a polypeptide marker No. 9;

No. 3, No. 5, No. 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 3, No. 5, No. 6 and No. 8 in combination with a polypeptide marker No. 9;

No. 3, No. 5, No. 7 and No. 8 in combination with a polypeptide marker No. 9;

No. 3, No. 6, No. 7 and No. 8 in combination with a polypeptide marker No. 9;

No. 4, No. 5, No. 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

No. 4, No. 5, No. 6 and No. 8 in combination with a polypeptide marker No. 9;

No. 4, No. 5, No. 7 and No. 8 in combination with a polypeptide marker No. 9;

No. 4, No. 6, No. 7 and No. 8 in combination with a polypeptide marker No. 9; or

No. 5, No. 6, No. 7 and No. 8 in combination with a polypeptide marker No. 9.

According to another preferred variant of the invention, the use of six polypeptide markers listed in table 1 or table 2 for the diagnosis of arteriosclerosis. In particular, this combination of polypeptide markers:

- № 1, № 2, № 3, № 4 and No. 5 in combination with one of the polypeptide markers No. 6, No. 7, No. 8 or No. 9;

- № 1, № 2, № 3, № 4 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

No. 1, No. 2, No. 3, No. 4 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 2, № 3, № 4 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 3, № 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

- № 1, № 2, № 3, № 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 2, № 3, № 5 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 3, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 2, № 3, № 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 3, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 4, № 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

- № 1, № 2, № 4, № 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 2, № 4, № 5 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 4, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 2, № 4, № 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 4, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 5, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 2, № 5, № 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 5, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 6, № 7 and No. 8 in combination : the AI with a polypeptide marker No. 9;

- № 1, № 3, № 4, № 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

- № 1, № 3, № 4, № 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 3, № 4, № 5 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 3, № 4, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 3, № 4, № 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 3, № 4, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 3, № 5, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 3, № 5, № 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 3, № 5, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 3, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 4, № 5, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 4, № 5, № 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 4, № 5, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 4, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 5, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 2, № 3, № 4, № 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

- № 2, № 3, № 4, № 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 2, № 3, № 4, № 5 and No. 8 in combination with polypeptidyl marker No. 9;

- № 2, № 3, № 4, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 2, № 3, № 4, № 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 2, № 3, № 4, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 2, № 3, № 5, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 2, № 3, № 5, № 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 2, № 3, № 5, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 2, № 3, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 2, № 4, № 5, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 2, № 4, № 5, № 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 2, № 4, № 5, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 2, № 4, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 2, № 5, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 3, № 4, № 5, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 3, № 4, № 5, № 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 3, № 4, № 5, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 3, № 4, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 3, № 5, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 4, № 5, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9.

According to yet another pre is respectful variant implementation of the invention uses seven polypeptide markers listed in table 1 or table 2 for the diagnosis of arteriosclerosis. In particular, this combination of polypeptide markers:

- № 1, № 2, № 3, № 4, № 5 and No. 6 in conjunction with one of the polypeptide markers No. 7, No. 8 or No. 9;

- № 1, № 2, № 3, № 4, № 5 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 2, № 3, № 4, № 5 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 3, № 4, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 2, № 3, № 4, № 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 3, № 4, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 3, № 5, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 2, № 3, № 5, № 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 3, № 5, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 3, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 4, № 5, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 2, № 4, № 5, № 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 4, № 5, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 4, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 2, № 5, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 3, № 4, № 5, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 1, № 3, �� 4, No. 5, No. 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 3, № 4, № 5, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 3, № 4, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 3, № 5, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 1, № 4, № 5, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 2, № 3, № 4, № 5, № 6 and No. 7 in combination with one of the polypeptide markers No. 8 or No. 9;

- № 2, № 3, № 4, № 5, № 6 and No. 8 in combination with a polypeptide marker No. 9;

- № 2, № 3, № 4, № 5, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 2, № 3, № 4, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 2, № 3, № 5, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9;

- № 2, № 4, № 5, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9; or

- № 3, № 4, № 5, № 6, № 7 and No. 8 in combination with a polypeptide marker No. 9.

According to another preferred variant of the invention using eight polypeptide markers listed in table 1 or table 2 for the diagnosis of arteriosclerosis. In particular, this combination of polypeptide markers:

- № 1, № 2, № 3, № 4, № 5, № 6, № 7 and No. 8;

- № 1, № 2, № 3, № 4, № 5, № 6, № 7 and # 9;

- № 1, № 2, № 3, № 4, № 5, № 6, № 8 and # 9;

- № 1, № 2, № 3, № 4, № 5, № 7, № 8 and # 9;

- № 1, № 2, № 3, № 4, № 6, № 7, № 8 and # 9;

- № 1, № 2, № 3, � 5, the # 6, # 7, # 8 and # 9;

- № 1, № 2, № 4, № 5, № 6, № 7, № 8 and # 9;

- № 1, № 3, № 4, № 5, № 6, № 7, № 8 and No. 9; or

- № 2, № 3, № 4, № 5, № 6, № 7, № 8 and # 9.

According to another preferred variant of the invention, the use nine polypeptide markers listed in table 1 or table 2 for the diagnosis of arteriosclerosis. In particular, this combination of polypeptide markers:

- № 1, № 2, № 3, № 4, № 5, № 6, № 7, № 8 and # 9.

In addition to the above polypeptide markers as a marker for diagnosis of arteriosclerosis can use the factor of the presence or absence of at least one other polypeptide when the polypeptide is indicative of arteriosclerosis or indicating the absence of arteriosclerosis (control, e.g., healthy state). The specified polypeptide, for example, consists of at least ten amino acids that are linked by peptide bonds. Preferably, the peptide consists of a maximum of 100 amino acids. Further, preferably the polypeptide has a molecular weight of from about 500 Da to 15,000 Da, even more preferably from about 750 Da to about 10,000 Da, in particular from about 1000 Da to about 7500 Yes. Thus, the markers can be chemically modified, for example, by post-translational modifications like glycosylation, phosphorylation, alquiler the existence or formation of disulfide bonds, or may be changed due to other reactions, for example, in the framework of the cleavage reaction. In addition, the polypeptide markers can also be chemically modified, for example oxidized, in the framework of the clean samples.

In the case of samples which determine the presence or absence of proposed according to the invention the polypeptide marker or proposed according to the invention the polypeptide markers, we can talk about any sample, which is obtained from the body of the individual. In the case of this sample can be a sample with the polypeptide composition which is suitable to receive testimony on the status of the individual (patient with arteriosclerosis or healthy, that is the individual without arteriosclerosis). For example, it may be blood, urine, synovial fluid, tissue fluid, the secret of the body, sweat, cerebrospinal fluid, lymph, intestinal juice, gastric juice, pancreatic juice, bile, tears, tissue sample, semen, vaginal fluid or sample of feces. Preferably the sample is a liquid sample.

According to a preferred variant implementation in the case of the sample it is the breakdown of urine or breakdown of the blood, the preferred sample is serum or blood plasma.

Urine sample is preferably selected in accordance with the prior art. Preferably, according to the present invention, as PR the urine would use the middle part of urine during urination. A urine sample may be selected, for example, by using external urinals, as described in International application WO-01/74275.

Blood samples can be extracted is known in the prior art methods, for example from a vein, artery or capillary. Usually a sample of blood is obtained by the fact that the individual take venous blood with a syringe, for example, from hands. The term "blood sample" also includes sample derived from the blood through further ways of purification and separation, such as blood plasma or serum.

The blood plasma obtained from the blood that removes cellular components, for example, by centrifugation. Centrifugation can be performed, for example, in the presence of anticoagulants, such as, for example, sodium citrate, as in the plasma are still coagulation factors. Serum essentially corresponds to the blood plasma, from which the removed active against clotting proteins - mainly fibrinogen, for example, by coagulation.

Methods of obtaining blood plasma and blood serum are well known in the prior art. In addition, in the International application WO-04/65958 describes how the removal of fibrinogen from plasma. Compositions for separating serum or plasma are described, for example, in International application WO-03/48764 (see also U.S. patent 2004/129631). Devices and methods for collection of samples plasmas is or serum from the blood describes, for example, in U.S. patent 2004/31746.

The presence or absence of a polypeptide marker in a sample can be determined by any known in the prior art method, which is suitable for determination of polypeptide markers. Specialist such methods are known. In principle, the presence or absence of a polypeptide marker can be determined by direct methods, such as mass spectrometry, or indirect ways, such as by using ligands.

If necessary or desirable, the sample taken from the individual, such as a urine sample or blood, before determining the presence or absence of the polypeptide marker or polypeptide markers can be subjected to pre-treatment by any suitable means, for example, to clean or separation. Processing may include, for example, purification, separation, dilution or concentration. The methods can be, for example, centrifugation, filtration, ultrafiltration, dialysis, precipitation or chromatographic methods, such as affinity separation or separation by ion-exchange chromatography, electrophoretic separation, i.e. separation due to the different nature of the migration of electrically charged particles in solution when applying the electric field. Specific examples include gel electrophoresis, two-dimensional, electropho the ez-polyacrylamide gel (2D-PAGE), capillary electrophoresis, metallofonda chromatography, immobilized metallofonda chromatography (IMAC), the affine chromatography based on lectins, liquid chromatography, high performance liquid chromatography (HPLC), normal and reversed-phase HPLC, cation exchange chromatography and selective binding to surfaces. All of these methods are well known to the specialist and the specialist can choose the method depending on the sample and method of determining the presence or absence of the polypeptide marker or polypeptide markers.

According to one variant of the invention, the sample, before its division, is subjected to separation by capillary electrophoresis, purified by ultracentrifugation and/or by ultrafiltration into fractions, which contain the polypeptide markers of a certain molecular size.

To determine the presence or absence of the polypeptide marker is preferably used mass spectrometric method, and this method can be predvklyuchen purification or separation of the sample. Mass spectrometric analysis, in comparison with common currently, methods, has the advantage that the concentration of many (>100) polypeptide in a sample can be determined by only the analysis is. You can use any type of mass spectrometer. Using mass spectrometry of complex mixtures can be defined generally 10 fmol polypeptide marker, therefore, 0.1 ng of protein with a mass of 10 kDa, with a measurement accuracy of approximately ±0,01%. In the case of mass spectrometers device for the formation of ions is associated with a suitable analyzer. For example, often use the interface blocks for ionizatsii electronic spray (ESI) for purposes of determining ions from liquid samples, while the laser matrix-desorption ionization (MALDI) is used primarily for producing ions from individually prepared samples. Commercially available various types of analyzers, such as analyzers of the type of ion trap or time-of-flight (TOF) analyzers. As ESI, and MALDI essentially can be combined with all types of mass spectrometers, although ESI is usually combined with ion traps, and combined with MALDI-TOF.

In the case of ionization by electron spray (ESI) in solution the molecules are sprayed, in particular, under the influence of high voltage (e.g., 1-8 kV), with the first form charged droplets, which due to evaporation of the solvent becomes less. Finally, the formation of free gaseous ions.

In the case of mass spectrometry using time-of-flight mass spectrometer TOF) make a certain accelerating voltage, which gives ions equally large kinetic energy. Then very accurately measure the time required appropriate ions to pass a certain drift section for passage of the pipe, since for the same kinetic energy the velocity of the ions depends on their mass. TOF Mass spectrometers have a very high scanning speed and thus achieve a good resolution.

Preferred methods for determining the presence or absence of polypeptide markers include gas-phase ion spectrometry, a mass spectrometry laser desorption/ionization, SELDI-TOF mass spectrometry (TOF mass spectrometry from surface-enhanced laser desorption/ionization), MALDI-TOF-mass spectrometry (time-of-flight mass spectrometry with laser matrix-desorption ionization), 2D-PAGE-mass spectrometry and capillary electrophoresis - mass spectrometry (CE-MS).

2D-PAGE is usually used for the separation of polypeptides and can be used in conjunction with mass spectrometry to identify individual polypeptides (2D-PAGE/MS). By 2D-PAGE, you can recognize more than 1000 protein sites. However, each individual polypeptide website needs to be identified separately using mass spectrometric method.

In SELDI system the most important component is the protein chip. EN lisaraye sample is applied directly on the matrix areas. There are different matrices, such as, for example, chromatographic matrix, which are hydrophobic, hydrophilic, cation-exchange, anion-exchange and immobilized surface with affinity to metal ions, and productiviprovincial matrix with chemical groups that allow you to be covalent binding. Preferred cation-exchange surface. In the case of SELDI-way only detect polypeptides that are actually in contact with the surface of the chip. After binding polypeptides samples use energy absorbent matrix at each site. After crystallization of the matrix a set of protein chip matrix estimate, as a rule, used for the analysis of the reader of the protein chip.

The reader is TOF(time-of-flight)mass spectrometer, in which proteins desorbers and ionized with a laser. As subjected to the crystallization of proteins is equally distributed on the surface area, the ionizing laser beam always refers to a representative cross-section of molecules in an analyte, which allows quantitative determination. After ionization polypeptides throw the acceleration in the electric field before they enter the detector. The duration of the touch laser irradiation surface of the die the flesh until the molecules in the detector allows precise determination of the molecular mass of the polypeptide in the sample (see, for example, following a review paper Merchant M. Weinberger S.R., Electrophoresis,212, 1164-1177 (2000)).

A particularly preferred method is the CE-MS, in which capillary electrophoresis is associated with mass spectrometry. This method is described in detail, for example, in the patent application Germany 10021737, articles Kaiser and others (J. Chromatogr. And,1013, 157-171 (2003), as well as Electrophoresis,25, 2044-2055 (2004)) and in article Wittke and others (Journal of Chromatography A,1013, 173-181 (2003)). The CE-MS Method allows high selectivity to determine the presence of hundreds of polypeptide markers simultaneously in a short time and in limited quantity. After polipeptidnaja the sample studied, prepared sample measured polypeptide marker and it can be compared with the sample of the patient, respectively, of a healthy person. In most cases the diagnosis of arthritis is sufficient to use only one or a limited number of polypeptide markers. In particular, one can define 1, 2, 3, 4, 5, 6, 7, 8 or 9 polypeptide markers listed in table 1 or 2, preferably in the above combinations of polypeptide markers. Hereinafter, preferred CE-MS method, which includes capillary electrophoresis, associated, for example, in-line with ESI-TOF analyzer. In this case, the sample contribute capillary electrophoresis and applied voltage, for example, up to 50 kV, about what bennoti up to 30 kV.

For CE-MS is preferable to use of volatile solvents and it is best to work in essentially salt-free conditions. Examples of such solvents include acetonitrile, isopropanol, methanol and the like. The solvents can be mixed with water or a weak acid (for example, of 0.1% formic acid) for protonation of the analyte. Polypeptide markers in a sample are separated according to their size and charge, which specifies the time span in the capillary.

Using capillary electrophoresis molecules can be separated according to their charge and size. While supplying the current of the neutral particles migrate with the velocity of electroosmotic flow, the cations are accelerated towards the cathode and anions to slow down. The advantage of the capillary during electrophoresis is favorable ratio of surface to volume, which makes possible a good diversion released when current passes through dzhoulevo heat. This, in turn, allows you to attach a high voltage (typically up to 30 kV) and at the same time to achieve high separation capacity and short time of analysis.

In the case of capillary electrophoresis, as a rule, use a capillary made of quartz glass. By using capillaries with narrow width to avoid light as radial diffusion, and convection. So that, when the applied voltage is the situation occurring in the capillary due to the passage of current heat was enough to take in the environment, the inner diameters of the capillaries should be small. Usually, the capillaries with internal diameters of 50 to 75 microns. Used lengths are, for example, 30-100 see In the case of capillaries with these dimensions, as a rule, provided that the temperature rise in the capillary due to the escaping ion heat does not lead to the formation of gas bubbles in the capillary. Moreover, the separation capillaries, as a rule, consist of a plastic shell of quartz glass. The capillaries can be raw, that is, on the inner side have their inherent hydrophobic groups, and also on the inner side can be coated. The hydrophobic coating can be used to improve the degree of separation. In addition to the voltage can be applied also to the pressure that is normally in the range 0-1 lb/in2. Pressure also can be applied only at the time of application or the time to change.

To improve the degree of separation during boot, you can use the Protocol "stacking": before loading the sample base load, then test, then acid. Thus, ions of the analyte are between acid and base. When applied voltage, the positively charged analyte ions move towards the base. That is, they become negatively charged and move in the opposite direction, where they are charged positively. This is repeated until the neutralization of acids and bases. So get a concentrated sample. Before loading the sample can be diluted with a suitable buffer.

For definitions separated by capillary electrophoresis polypeptide marker device for capillary electrophoresis is preferably associated with a mass spectrometer. Mass spectrometry allows the determination of molecular masses of free ions in a high vacuum. Mass spectrometer contains a mass analyzer, which separates the ions according to their ratio of mass-to-charge (m/z), and a detector.

According to another preferred method for determining polypeptide markers in a sample polypeptide markers of the sample separated by capillary electrophoresis, and then directly ionize and to determine translate into online through the interface block into the associated mass spectrometer.

Another object of the present invention is a method of diagnosis of arteriosclerosis:

a) determining the presence or absence of at least one polypeptide marker in a sample of the individual, and the polypeptide marker is defined as in paragraph 1 or 2 of the claims, and

b) establishing the probability of the presence of atherosclerosis in an individual, and

i) when verojatnost the presence of the polypeptide marker in a patient individual higher than the probability of the presence of this polypeptide marker in the control of the individual, then the presence of the polypeptide marker indicates a higher probability of presence of arteriosclerosis;

ii) when the probability of the presence of the polypeptide marker in a patient of an individual is lower than the probability of the presence of the polypeptide marker in the control of the individual, then the absence of a polypeptide marker indicates a higher probability of presence of arteriosclerosis;

iii) when the probability of the presence of the polypeptide marker in a patient of an individual is higher than the probability of the presence of this polypeptide marker in the control of the individual, then the absence of a polypeptide marker indicates a higher probability of the absence of arteriosclerosis; or

iv) when the probability of the presence of the polypeptide marker in a patient of an individual is lower than the probability of the presence of the polypeptide marker in the control of the individual, then the presence of the polypeptide marker indicates a higher probability of the absence of arteriosclerosis.

According to one option proposed in the invention method, the probability in the case of stage b) in relation to the presence of the polypeptide marker in a patient with arteriosclerosis of the individual (patients), respectively, in the control of the individual are the two which are as follows:

Polypeptide marker No.Molecular weight [Yes]The migration time [min]The probability of the presence of the polypeptide marker
the patient individualthe control of the individual
12511,826,40,600,00
21864,749,20,200,78
32799,846,00,200,78
41340,666,70,100,67
51422,861,20,000,56
65882,033,30,00 0,56
71397,424,90,500,00
81886,644,70,500,00
94642,637,50,500,00

According to a preferred variant of controlling the individual is not sick arteriosclerosis individual. According to still another preferred variant implementation, he is a man without arteriosclerosis.

According to the proposed invention, the method preferably can be used for the diagnosis of multiple polypeptide markers. In particular, one can use at least 2, 3, 4, 5, 6, 7, 8 or 9 polypeptide markers listed in table 1 or table 2. Further, more preferably, in the case of combinations of polypeptide markers are a combination of the above.

To determine the probability of the presence of arteriosclerosis when using multiple markers specialist can use a well-known statistical methods. For example, you can use described Weissinger and others (Kidey Int., 65(6), 2426-24 (2004, June) method (statistical Forests-way) using a computer program, such as S-Plus.

According to the proposed invention the presence of the polypeptide markers № 1, № 7, № 8 and/or # 9 in the sample and/or absence of polypeptide markers№ 2, № 3, № 4, № 5 and/or No. 6 in the sample indicates the presence of arteriosclerosis.

To determine the presence or absence in the case of proposed according to the invention method, you can use the above cleaning methods, separation and detection. To detect the presence or absence of the polypeptide marker or polypeptide markers preferably using capillary electrophoresis, gas-phase ion spectrometry, and/or mass spectrometry.

According to a particularly preferred variant implementation prior to measurement of molecular weight polypeptide markers perform capillary electrophoresis and/or use mass spectrometry to detect the presence or absence of the polypeptide marker or polypeptide markers.

Example

1. Sample preparation

To determine the polypeptide markers for the detection of arteriosclerosis used the plasma. Plasma was obtained from healthy donors (control)and patients required dialysis who suffer from arteriosclerosis. For the enterprises of plasma from healthy donors was used S-Monovetten with heparin (Sarstedt, Nürnbrecht, Germany), and subjected to dialysis patients was selected plasma, because they were treated with heparin. Heparin is used to prevent blood clotting and proteolytic activity and yet also for the protection contained in the plasma proteins and peptides from splitting enzymes.

For subsequent CE-MS measurements available in the plasma at very high concentrations of proteins as albumin and immunoglobulins, had to be separated by ultrafiltration. For this selected 500 μl of plasma was mixed with 2 ml of buffer to filter (4 mol/l urea, 0.1 mol/l NaCl, 0.01% ammonia). These sample volumes, constituting 2.5 ml, was subjected to ultrafiltration (Centrisart 20 MWCO, Vivascience, Hannover, Germany). The ultrafiltration was carried out in a centrifuge at a speed of 3400 rpm up to obtain 2 ml of ultrafiltrate.

Received 2 ml of the filtrate was then introduced into the column Pharmacia C-2 (Pharmacia, Uppsala, Sweden) to remove urea, salts and other interfering components. Related polypeptides were then suirable from C-2-column with 50% acetonitrile and 0.5% formic acid in water and liofilizirovanny. For CE-MS measurements polypeptides then resuspendable in 20 µl of water (HPLC-purity, Merk, Darmstadt, Germany).

2. CE-MS-Dimension

CE-MS Measurement was performed using the system for capillary electrophoresis firm Beckman Coulter P/ACE MDQ System; Beckman Coulter Inc., Fullerton, CA, USA) the ESI-TOF mass spectrometer company Applied Biosystems (Mariner Biospectrometry Workstation; Applied Biosystems, Foster City, CA, USA).

CE Capillary was received from the company Beckman Coulter (see above), they had a ratio of ID/OD, equal 75/360 μm, and a length of 90 cm the Mobile phase for the CE separation consisted of 30% methanol and 0.5% formic acid in water, the same mixture used for the Sheath Flow in the mass spectrometer, in this case, with a flow rate of 2 µl/min Linking CE and MS was performed using the set of CE-ESI-MS Sprayer Kit (Agilent Technologies, Waldbronn, Germany).

For injection 1 µl of sample was applied a pressure of 1 lb/inch2the duration of injection was 20 seconds. When using these parameters in the capillary was injectively approximately 100 nl of the sample, this corresponds to approximately 0.25% of the volume of the capillary. For concentration of the sample in the capillary used a technology called "stacking". Thus, before injection of the sample for 7 seconds (at a pressure of 1 lb/in2) was injectively 1 mol/l NH3after injection of the sample for 5 seconds have introduced 2 mol/l solution of formic acid. When the application of the separation voltage (30 kV) analytes automatically centered between these solutions.

Following the CE separation was carried out according to the method of pressure: 40 minutes at a pressure of 0 lb/in2then 2 minutes at a pressure of 0.1 lb/in2, 2 minutes at a pressure of 0.2 lb/in2, 2 minutes at a pressure of 0.3 lb/in2, 2 minutes at a pressure of 0.4 lb/in2, ZAT is 32 minutes at a pressure of 0.5 lb/in 2. The total duration of the separation process was thus 80 minutes.

To get from MS if possible, good signal intensity was eliminated gas-spray. Attached to the needle-spray voltage to create electrospray was 3,200 In consequence of which the result was achieved using existing spray voltage across the capillary, about 27 kV. Other adjustments in the case of Mariner-mass spectrometer optimized according to the instructions of the manufacturer for the detection of the peptide. The spectra were taken in the mass region from m/z 400 to m/z 2500 and all accumulated within 3 seconds.

3. The standards for CE-dimension

For control and standardization CE measurements used the following proteins, respectively, polypeptides, which are listed CE migration times:

(it can be used to normalize the CE era peptide markers 1-9, as well as the measured samples)

the migration time of the protein/polypeptide

Aprotinin (SIGMA, Taufkirchen, Germany, registration number AI 153): 9,2 min

ribonuclease (SIGMA, Taufkirchen, Germany, registration number R4875): 10,9 min

secrete lysozyme (SIGMA, Taufkirchen, Germany, registration number L7651): 8,9 min

“REV”, sequence: REVQSKIGYGRQIIS: 15,6 min

“ELM”, sequence: ELMTGELPYSHINNRDQIIFMVGR: 23,4 min

“KINCON”, sequence: TGSLPYSHIGSRDQIIFMVGR: 20,0 min

“GIVLY”, a follower of the awn: GIVLYELMTGELPYSHIN: 36,8 minutes

Proteins/polypeptides used, respectively, at a concentration of 10 pmol/ál in water.

“REV”, “ELM”, “KINCON” and “GIVLY” are synthetic peptides.

Molecular weight peptides, and also found according to the mass spectrum of mass of individual charged States are as follows:

1. Method of diagnosis of arteriosclerosis, which includes stages:
a) determining the presence or absence of at least one polypeptide marker in a sample of the individual, and the polypeptide marker selected from the polypeptide markers№ 1, № 2, № 3, № 4, № 5, № 6, № 7, № 8 or No. 9, which are characterized by the following values of molecular mass:

Polypeptide marker No.Molecular weight (Da)
12511,8
21864,7
32799,8
41340,6
51422,8
65882,0
71397,4
81886,6/td>
94642,6

and
b) establishing the probability of the presence of atherosclerosis in an individual, and
i) when the probability of the presence of the polypeptide marker in a patient of an individual is higher than the probability of the presence of this polypeptide marker in the control of the individual, then the presence of the polypeptide marker indicates a higher probability of presence of arteriosclerosis;
ii) when the probability of the presence of the polypeptide marker in a patient of an individual is lower than the probability of the presence of the polypeptide marker in the control of the individual, then the absence of a polypeptide marker indicates a higher probability of presence of arteriosclerosis;
iii) when the probability of the presence of the polypeptide marker in a patient of an individual is higher than the probability of the presence of this polypeptide marker in the control of the individual, then the absence of a polypeptide marker indicates a higher probability of the absence of arteriosclerosis; or
iv) when the probability of the presence of the polypeptide marker in a patient of an individual is lower than the probability of the presence of the polypeptide marker in the control of the individual, then the presence of the polypeptide marker indicates a higher probability of the absence of arteriosclerosis.

2. The method according to claim 1, where polypeptidyl marker selected from the polypeptide markers No. 1, № 2, № 3, № 4, № 5, № 6, № 7, № 8 or No. 9, which are characterized by the following values of molecular weight and time values migration:

Polypeptide marker No.Molecular weight (Da)The migration time (min)
12511,826,4
21864,749,2
32799,846,0
41340,666,7
51422,861,2
65882,033,3
71397,424,9
81886,644,7
94642,637,5

3. The method according to claim 1 or 2, where the individual probabilities in the case of stage b) are the following:

Polypeptide marker No.The probability of the presence of the polypeptide marker
the patient individualthe control of the individual
10,600,00
20,200,78
30,200,78
40,100,67
50,000,56
60,000,56
70,500,00
80,500,00
90,500,00

4. The method according to claim 1 or 2, where using at least 2, 3, 4, 5, 6, 7, 8 or 9 polypeptide markers.

5. The method according to claim 1 or 2, which use a combination of at least two polypeptide markers, in particular polypeptide markers:
№ 1 and № 2, № 1 and № 3, № 1 and № 4, № 1 and № 5, № 1 is No. 6, № 1 and № 7, № 1 and № 8, № 1 and № 9;
№ 2 and № 3, № 2 and № 4, № 2 and № 5, № 2 and № 6, № 2 and № 7, № 2 and № 8, № 2 and № 9;
№ 3 and № 4, № 3 and № 5, № 3, № 6, № 3 and № 7, № 3 and № 8, № 3 and № 9;
№ 4 and № 5, № 4 and № 6, № 4 and № 7, № 4 and № 8, № 4 and № 9;
№ 5 and № 6, № 5 and № 7, № 5 and № 8, № 5 and № 9;
# 6 and # 7, # 6 and # 8, # 6 and # 9;
No. 7 and No. 8, No. 7 and No. 9; or
# 8 and # 9;
and moreover, polypeptide markers defined in claim 1 or 2.

6. The method according to claim 1, where using at least one other polypeptide as a marker.

7. The method according to claim 1, where the migration time is determined by capillary electrophoresis at 30 kV using a mobile phase of 30% vol. methanol and 0.5% vol. formic acid in water in the capillary length 90 cm

8. The method according to claim 1, where the sample of the individual is a urine sample or a blood sample, particularly a sample of serum or plasma sample.

9. The method according to claim 1, where the presence of polypeptide markers № 1, № 7, № 8 and/or # 9 indicates the presence of arteriosclerosis.

10. The method according to claim 1, where the absence of polypeptide markers№ 2, № 3, № 4, № 5 and/or No. 6 indicates the presence of arteriosclerosis.

11. The method according to claim 1, where detecting the presence or absence of the polypeptide marker or polypeptide markers using capillary electrophoresis, gas-phase ion spectrometry, and/or mass spectrometry.

12. The method according to claim 1, where prior to the determination of molecular weight polypeptide m is rcrow perform capillary electrophoresis.

13. The method according to claim 1, where detecting the presence or absence of the polypeptide marker or polypeptide markers using mass spectrometry.



 

Same patents:

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to leprology, and can be used particularly for the detection of specific liver disease in the leprosy patients. Blood serum of the leprosy patients is analysed to determine the level of US-M.leprae and DIS-BSA leprosy mycobacteria antigen antibodies, and also the concentration of alpha-1-antitrypsin. If observing the level of antibodies to two antigens increased as compared to a norm and the concentration of alpha-1-antitrypsin increased higher than 346.7 mg/dl, liver disease is detected in the leprosy patients.

EFFECT: method provides higher accuracy.

4 ex

FIELD: medicine.

SUBSTANCE: essence of invention includes selection of sample for testing from area of malignant affection in patient, suffering from cancer of large intestine, selection of control sample, measurement of the level of one or several genetic markers, selected from the group, comparison of measured levels in experimental samples with control ones. Reducing of levels of one or several genetic markers in comparison with control sample, indicates increased resistance to docetaxel. Also described are sets which allow to predict or realise monitoring of patient's response to docetaxel.

EFFECT: identification of genetic markers, by which it is possible to realise patient's response to chemical therapy by docetaxel.

18 cl, 9 ex, 3 tbl, 16 dwg

FIELD: chemistry; biochemistry.

SUBSTANCE: present invention relates to molecular biology and can be used in designing agent and methods of modulating body functions associated with HGF/c-met signalling pathway. The invention discloses HGF/c-met polypeptide-antagonists which are mutant forms of HGF which contain a mutation in the N-terminal part of the β-chain and/or in its dimerisation part. The disclosed polypeptides have lower biological activity compared to wild type HGG and can be used in modulating activity of c-met, cell proliferation, cell migration and angiogenic cell activity.

EFFECT: invention describes a method of obtaining HGF muteins using DNA recombinant technology and agents which are necessary for its existence.

22 cl, 8 dwg, 1 ex

FIELD: medicine.

SUBSTANCE: in order to predict insufficiency of saturation with oxygen of peripheral blood of pregnant women who have herpes-virus infection, content of glucose-6-phosphat-dehydrogenase and TNFα in peripheral blood is determined. Discriminant equation D=-0.589·G-6-PDG+{+1.23·TNFα} is solved. If value of discriminant function is equal or greater than 89.95, predicted is development of oxygen insufficiency, accompanied with reduction in peripheral blood of pregnant women of pO2 with titre of antibodies to Herpes simplex virus 1:12800 to 27.63±0.7 mm Hg, and of HbO2 to 90.2±0.47% (control: pO2 - 39.22±0.5 mm Hg; HbO2 - 95.3±0.27%).

EFFECT: increased accuracy of determining possibility of development of oxygen insufficiency in pregnant woman with herpes-virus infection.

3 tbl

FIELD: medicine.

SUBSTANCE: in estimation of local inflammation activity in case of background diseases of neck of uterus sampling of analysed biomaterial is performed by bringing of device for native material removal to external neck of uterus fauces. Said device has elongated holder in form of rod with length, exceeding length of vagina. From distal side of rod, at angle to it, located is operation element in form of roller. Proximal end of rod serves as manipulator-handle. Sampled material is placed until complete submergence into tightly closable test tube, filled with 1.0 ml of 0.155 M solution of sodium chloride. Mixture is centrifuged for 10 minutes. Obtained materials are analysed by method of solid phase ELISA analysis on boards by means of set of reagents "Vector-Best". Concentration of cytokine of interleukin-6 in analysed samples is determined spectrophotometrically. If value is higher than 50 pg/ml to 92 pg/ml, conclusion about presence of chronic inflammation process is made. If value is higher than 92 pg/ml, conclusion about active course of inflammatory process and expressed immune response is made.

EFFECT: application of method allows to increase quality of analysed native material and accuracy of obtained result without complicating analysis process.

2 cl, 3 ex

FIELD: medicine.

SUBSTANCE: invention can be used for diagnostics of presence of destructive process in uterine appendages in case of localised peritonitis if gynecological genesis, and for selection of optimal treatment tactics. In order to realise the method content of C-reactive protein (CRP) in blood serum is determined by turbodimetric method, if values of C-reactive protein content are 130 mg/l and higher, purulent-destructive process in uterine appendages in case of localised peritonitis is diagnosed, if values of C-reactive protein content are lower than 130 mg/l diagnosed is purulent salpingitis in case of pelvioperitonitis.

EFFECT: application of method allows to detect presence of purulent-destructive process in uterine appendages in case of pelvioperitonitis in urgent way, which determines tactics of further treatment.

2 ex

FIELD: medicine.

SUBSTANCE: invention can be used for obtaining diagnostic and/or therapeutic agents, in particular antibodies, which specifically interact with proteins of cell surface of target cells. Claimed is method of screening phage display library and including it method of isolation of library element, which is specific partner of binding of target cell surface protein, and method of isolation of unknown cell surface protein, specifically interacting with library element. Method of screening phage display library according to invention includes contact of said library with cells of interest with further separation of cells, which bound with one or several elements of expression library, from non-bound elements by separation through organic phase, and differs by introduction of additional stage, which consists in carrying out before said separation of at least one stage of material washing, and ensures essential increase of method efficiency.

EFFECT: higher efficiency.

30 cl, 7 dwg, 6 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to ophthalmology, can be used for prediction of progression of myopia acquired in schools in 10-14 and 15-17 year old children. Blood serum is analysed for cartilage glycoprotein-39 with the use of sandwich-type enzyme immunoassay. If its concentration is within 34.7-53.4 ng/ml in 10-14-year-old children and 22-56.3 ng/ml in 15-17-year-old children, a permanent course of myopia is predicted. The concentration 18.5-33.9 ng/ml in 10-14-year-old children and 53.7-69 ng/ml in 15-17-year-old children enables to predict a progressive course of myopia.

EFFECT: use of the invention improves accuracy of the prediction of clinical course.

4 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to diagnostic techniques and concerns a diagnostic techniques technique for respiratory function of peripheral blood erythrocytes in pregnant women in herpes virus infection episode in the third trimester. The technique consists in determination of glutathione reductase and herpes virus antibody titre concentration by the enzyme immunoassay in peripheral blood in pregnant women suffered from a herpes virus infection episode. If glutathione reductase concentration is 8.36±0.137 E/g Hb (ref. - 7.82±0.185 E/g Hb), herpes virus antibody titre is 1:6400. If herpes virus antibody titre is increased to 1:12800, erythrocyte glutathione reductase concentration is decreased 4.48±0.22 E/g Hb.

EFFECT: technique is characterised by high sensitivity and allows predicting the development of impaired respiratory activity of erythrocytes in the pregnant women with the herpes virus infection episode.

FIELD: medicine.

SUBSTANCE: simultaneously two parametres are determined: for men - in ejaculate, and for women - in discharge of cervical canal, namely, slgA and lactoferrin. If level of secretory immunoglobulin A is 3.01±0.50 mcg/ml and lower and simultaneous level of lactoferrin is 6876±89 ng/ml and higher, gonococcus infection with systemic manifestations is diagnosed, if said condition is absent, localised gonorrhea is diagnosed.

EFFECT: application of claimed method allows carrying out differential diagnostics of various forms of gonococcus infection in case of its oligosymptomatic course.

3 ex, 2 tbl

FIELD: medicine, ophthalmology.

SUBSTANCE: in lacrimal liquid one should detect the content of interleukin 8 (IL-8) and that of interleukin 1 beta (IL-1β) to calculate prognostic coefficient (PC) due to dividing the first value by the second one by the following formula: At PC value being below 10.0 one should predict favorable disease flow, and at PC value being above 10.0 - unfavorable flow.

EFFECT: higher accuracy of prediction.

2 ex

FIELD: forensic medicine.

SUBSTANCE: for the purpose to detect the sequence of applied lesions at availability of several wounds, scratches and ecchymoses on a cadaver one should study the activity of alkaline peptides isolated out of affected tissue by the impact of blood neutrophils of healthy donors upon phagocytosis. Moreover, the highest stimulating effect belongs to the peptides isolated out of the lesion applied earlier. The method enables to detect the sequence of applied lesions more accurately and differentiate the repeated lesion applied 5 min later, or more.

EFFECT: higher efficiency and accuracy of detection.

2 ex, 2 tbl

FIELD: medicine, biochemistry.

SUBSTANCE: in blood serum one should detect the level of lactoferrin and biliary acids. At their ratio being equal to 5-17 it is necessary to detect chronic hepatitis of high activity.

EFFECT: higher accuracy of detection.

3 ex

FIELD: medicine.

SUBSTANCE: method involves determining cathepsin D activity in endometrium bioptate. The value being equal to or less than 0.1 units of enzymatic activity per hour, external genital endometriosis is diagnosed.

EFFECT: high accuracy of diagnosis.

1 tbl

FIELD: medicine.

SUBSTANCE: method involves studying lactoferrin content in blood serum and peritoneal exudates in postoperative period every day during the first three days. Lactoferrin concentration in blood serum being concurrently reduced by 0.02 mcmole/l or less and increasing lactoferrin concentration in peritoneal exudates by 0.04 mcmole/l or more, enteric detoxication is considered to be effective.

EFFECT: high quality of estimation.

2 tbl

FIELD: medicine.

SUBSTANCE: method involves determining plasminogen/plasmin, α2-macro-globulin, α1-antitripsin content at the first, third, fifth and tenth day. The plasminogen/plasmin level being equal to 66-74 mcmole/l or 100-120 mcmole/l, α2-macro-globulin level of 2.7-3.0 mcmole/l, α1-antitripsin content of 2.38-3.2 mcmole/l, systemic inflammatory response to purulent infection, light severity degree endotoxicosis is diagnosed and favorable disease outcome is predicted. The plasminogen/plasmin level being equal to 50-65 mcmole/l or 125-160 mcmole/l, α2-macro-globulin level of 2.3-2.6 mcmole/l, α1-antitripsin content of 3.3-4.0 mcmole/l, sepsis with organ and system dysfunction, moderate severity degree endotoxicosis is diagnosed and septic complication availability and lingering disease development course is predicted. The plasminogen/plasmin level being equal to 39-40 mcmole/l, α2-macro-globulin level of 1.58-2.08 mcmole/l, α1-antitripsin content of 5.0-6.2 mcmole/l, severe sepsis, septic shock, severe degree endotoxicosis is diagnosed and unfavorable disease outcome is predicted.

EFFECT: high accuracy of diagnosis.

5 tbl

FIELD: medicine, biochemistry.

SUBSTANCE: at testing one should precipitate high-molecular compounds with acetonitrile and register supernatant's spectral characteristics. Supernatant should be applied onto a paper filter, dried and put into solution containing aromatic aldehyde, acetone and concentrated hydrochloric acid taken at weight ratio of 70:5:1 to be kept for 2-3 min. Then it should be once again dried up to detect qualitative and semiquantitative content of oxidized tryptophan metabolites by intensity and chromatic shades. Moreover, by chromatic shades of yellow dyeing it is possible to detect the content of hydroxylated metabolites and by chromatic shades of violet dyeing - that of unhydroxylated ones.

EFFECT: higher significance of detection.

3 ex

FIELD: medicine, anesthesiology, resuscitation.

SUBSTANCE: in patients one should study the content of lactoferrin in peritoneal exudates during the 1st d of postoperational period and at decreased value being below 3500 ng/ml on should predict unfavorable result. The suggested method provides correction of possible postoperational complications that deteriorate the flow of peritonitis and lead to lethal result.

EFFECT: higher accuracy of prediction.

3 ex

FIELD: veterinary medicine.

SUBSTANCE: method involves determining low and middle molecular mass substances content in blood plasma and erythrocytes and general blood plasma albumin concentration. Integral index is calculated on basis of obtained values using formula II=100*S238-298(plasma)/S238-298(erythrocytes)*GAC, where S238-298(plasma) and S238-298(erythrocytes) are the low and middle molecular mass substances content in blood plasma and erythrocytes, respectively, determined from area of figures restricted by spectral curves in wavelength range of 238-298 nm and abscissa axis (conditional units2); GAC is the general blood plasma albumin concentration (g/l). The value being from 2.1 to 3.0, the first endotoxicosis degree is diagnosed. The value being from 3.1 to 4.5, the second endotoxicosis degree is diagnosed. The value being from 4.5 to 6.0, the third endotoxicosis degree is diagnosed. The value being greater than 6.0, the fourth endotoxicosis degree is diagnosed. The normal value is equal to 0.5-2.0.

EFFECT: high accuracy of diagnosis.

1 dwg, 1 tbl

FIELD: medicine.

SUBSTANCE: method involves separating blood serum proteins into fractions, determining albumins and alpha-2-globulins content and controlling their content changes during the disease development process. Gamma-globulin content is determined in per cent ratio with respect to total protein quantity. Then, changes in the fractions content are controlled from the first to the third week. Albumin content being in norm and alpha-2-globulins content becoming greater to the end of the first week by 30-50% when compared to normal value and dropping to norm at the second week end and gamma-globulin content increasing from norm by 10-30% to the second or the third week, high inflammatory process activity is to be diagnosed. Albumin content dropping by 10-30% from normal value at the second week, alpha-2-globulins content growing by 10-20% of norm and gamma-globulin content dropping by 30-50% at the second or the third week when compared to norm, low inflammatory process activity is to be diagnosed.

EFFECT: high accuracy and reliability of diagnosis.

Up!