Method for diagnosis of internal endometriosis

FIELD: medicine, gynecology.

SUBSTANCE: invention relates to a method for diagnosis of internal endometriosis in peripheral venous blood of women wherein the relative content of lymphocytes CD25+ is determined. Internal endometriosis is diagnosed at values of this index 6% or above. Proposed method provides carrying out diagnosis of internal endometriosis in women with high precision, sensitivity and specificity that allows carrying out the correct and well-timed necessary complex of curative-prophylactic treatment.

EFFECT: improved method for diagnosis.

1 tbl, 3 ex

 

The invention relates to medicine, namely to gynecology, and can be used to diagnose internal endometriosis.

The relevance of the proposed method is determined by a high prevalence (from 4 to 17% of women of reproductive age) genital endometriosis, in the structure of the disease, the frequency of the internal endometriosis is 41.5% [1]. The internal diagnosis of endometriosis is based on the results of invasive traumatic instrumental study (hysteroscopy, hysterosalpingography) or on the basis of histological examination remote uterus. In this regard, the development of non-traumatic reliable laboratory diagnostic criteria of endometriosis is of great importance for practical health care.

A known method for the diagnosis of internal endometriosis, based on the definition in the peripheral blood of women phosphatidylinositol (PHI), PHI values within 391,6±17.5 mmol per 1 liter of blood is diagnosed endometriosis of the uterus [3].

The disadvantages of the method:

1) is similar to the decline PHI observed in many diseases, including those associated with proliferative processes, or changes in the properties of cell membranes, which reduces the informativeness of the way;

2) the method is used only in combination with other methods is iagnostic internal endometriosis (ultrasound, hysteroscopy with histological examination, etc. and may not be used as a self;

3) time-consuming and costly;

4) do not specify the precision, sensitivity and specificity of the method.

The closest technical solution to the claimed method is a method for diagnosing an internal endometriosis, based on biochemical analysis of blood serum, characterized in that the serum to determine the level of acid phosphatase and identifying its values 50,91±6.95 mmol/l diagnose internal endometriosis of the uterus [4].

This method was chosen as the closest analogue (prototype). However, it has several drawbacks:

1) the enzyme is not resistant, rapidly disintegrates;

2) the level of acid phosphatase depends on somatic pathology;

3) the authors do not give an interval in which the change rates of acid phosphatase in this pathology;

4) do not specify the precision, sensitivity and specificity of the method.

These drawbacks are proposed to be eliminated in the present method.

The claimed technical result is achieved by the fact that in the peripheral venous blood of women determine the relative content of CD25+ lymphocytes and the values of this index equal to or greater 6%, diagnosed endometriosis of the uterus with an accuracy of 86%, feelings is of 92%, the specificity of 83%.

The novelty of the proposed method lies in the fact that for the first time proposed to diagnose internal endometriosis, by definition, the relative content of CD25+ lymphocytes in the peripheral blood.

Previously the content of CD25+ lymphocytes used in the diagnosis of acute viral infection [6], autoimmune diseases [2] and the prediction of postoperative infectious complications [5]. For the diagnosis of internal endometriosis this indicator is used for the first time.

It is known that CD25+ molecule, representing α the chain of the receptor for interleukin 2 (IL-2), is expressed on the surface membrane of lymphocytes in the early stages of activation [7]. High content in peripheral blood of women with an internal endometriosis activated CD25+ lymphocytes, in our opinion, may be associated with the development of systemic immune response is present in an organism modified autologous endometrial cells.

The method is standard, as follows.

1) Isolation of lymphocytes from peripheral venous blood is carried out by the standard method [8];

2) 100 μl of lymphocytes in a concentration of 1×106cells/ml are incubated with 20 μl of monoclonal anti-A labeled with FITC, (OOO "Medispectra", Moscow, Russia, registration certificate No. 29/24070700/2013-01 14 and the nya 2001) for 30 minutes at room temperature in the dark, then the cells washed 1 time with 1 ml of saline by centrifugation at 1500 rpm for 2 minutes. Cells resuspended in 500 μl of saline. The reaction accounting is performed on a flow cytometer, determining the content of CD25+ cells in the lymphocyte gate.

The use of the proposed method with high accuracy of 86%, sensitivity 92%, specificity of 83% to diagnose internal endometriosis.

Distinctive features of the method are:

set diagnostic parameter of the relative content of CD25+ lymphocytes in the peripheral blood of women in the quantitative values of which 6% or more internal endometriosis is diagnosed.

The essence of the proposed method is illustrated by the following examples.

Example 1. Patient P. 40 years. Asked about pain in lower abdomen, occasional spotting after menstruation. Sick for about 6 years. History 2 timely delivery. Conducted repeated antibiotic treatment both outpatient and inpatient short-term effect on chronic 2-party-oophoritis, endomyometritis. Introduced IUD is removed after 1.5 years about the exacerbation of the inflammatory process (pain, krivomazova).

At vaginal examination: increased uterine minor to 5 weeks pregnant the minute, tictacti in the vaults.

Clinical diagnosis: chronic endomyometritis, bilateral salpingo-oophoritis.

Data ultrasound examination: the body of the uterus with smooth contour, mesh heterogeneous structure, high ahopelto, size 64×40×54 mm, M-echo - 12 mm, cervix 42×38×39 mm, homogeneous structure. The right ovary in the intestinal loops with fuzzy circuit 37×23 mm with exonerative inclusions on the periphery with a diameter of 7×10×13 mm Left ovary is not defined. Conclusion: the pattern corresponds to endomyometritis, oophoritis.

According to immunological examination by the present method, the relative content of CD25+ lymphocytes in the peripheral blood of 10.7%, which is higher than the parameter for the internal endometriosis.

Conclusion: internal endometriosis is diagnosed.

The treatment scheme used for endometriosis, treatment of progestogen within 6 months, then were assigned to combined estrogen-gestagennye drugs. The pain disappeared with the beginning of the use of gestagens, normal menstrual cycle. By ultrasound: the body of the uterus with smooth contour of a homogeneous structure, the usual ahopelto, size 61×40×52 mm, M-echo - 7 mm cervix and ovaries without the speakers.

Improving clinical and the instrumental record of performance proves the correctness of the diagnosis and the treatment.

Example 2. Patient C. for 36 years. Appealed with complaints about krivomazova before and after menstruation, lower abdominal pain and lower back, more abundant menstruation last 3 cycles. In history one delivery and 2 medical abortion.

At objective inspection observed an increase in the uterus up to 5-6 weeks of pregnancy, the uterus is sensitive to palpation, is limited in mobility. The appendages are not defined, codes free, mucous allocation moderate.

Clinical diagnosis: internal endometriosis.

Data ultrasound: uterus size 62×54×68 mm inhomogeneous structure, with indistinct outline, the right ovary - 34×25 mm, left ovary - 35×23 mm, their structure and localization is not altered.

Conclusion: internal endometriosis.

According to immunological examination by the present method the indicator of the relative content of CD25+ lymphocytes in the peripheral blood of 3.8%, which is less than the stated parameter on the internal endometriosis.

Conclusion: internal endometriosis cannot be diagnosed. Obviously, there is endomyometritis.

Conducted anti-inflammatory treatment was effective: no pain in the lower abdomen and lower back, chromazone before and after menstruation, hyperprolinemia.

The positive effect of the course confirms the correctness of the diagnosis by Appl is the reception method.

Example 3. Patient A. 34 years. He complained of periodic abdominal pain, scanty bleeding from the genital tract within a few days after menstruation. History of timely delivery, 2 medical abortion.

Objective examination: the body of the uterus a little more normal size, heterogeneous structure, sensitive to palpation. In the field of appendages is determined by tictacti. Clinical diagnosis: endomyometritis.

The data of ultrasound scanning: the body of the uterus rounded, with indistinct outline of heterogeneous structure, size 61×44×64 mm M - echo - 10 mm Right ovary - 31×25 mm Left ovary - 30×26 mm with a dense center and follicular inclusions on the periphery of up to 11 mm, without dominant follicles. On the basis of ULTRASONIC research we can assume endomyometrial? or internal endometriosis?

According to immunological examination by the present method the indicator of the relative content of CD25%+ lymphocytes in the peripheral blood is 6.0%, which corresponds to the declared parameter. Conclusion: internal endometriosis is diagnosed. Held hormone therapy endometriosis. The patient noted improvement in health, no pain, normalization of the menstrual cycle, which indicates the correct diagnosis and treatment is.

Table 1
IndexThe number of the examined
Total female37
true-positive result12
false-positive result4
a true-negative result20
a false-negative result1

Total:

the accuracy of the proposed method with 86% sensitivity of the proposed method is 92% specificity of the proposed method is 83%

The advantages of the proposed method:

1. Can be used as a separate or diagnostic screening method.

2. Retraumatized.

3. High accuracy of 86%, sensitivity 92%, specificity 83%.

4. The duration of the method of determination of CD25+ lymphocytes does not exceed 1 hour and 30 minutes.

References

1. L.V. Adamyan, Kulakov V.I. Endometriosis: a Guide for physicians. - M.: Medicine. - Moscow, 1998.- 320 S.

2. Glazunova T.V., Pavlova IE, Rozanov, PU and other Immunological parameters in patients with concomitant autoimmune pathology /Medical immunology.- 2001.-V.3.- No. 2.- Pp.178-179.

3. V.V. Kulakov, Bakulev, L.P., Damirov M.M., Sarkisov SE, Slusar NN. To the question of diagnosis is the IR internal endometriosis/Akush. and gin.-1995.-№1.-Ñ.38-40.

4. Peresada O.A. method for the diagnosis of internal endometriosis. Patent No. 211729 Invention, 1998, 22, S).

5. Polosukhina ER, Zabotina T.N., Perlin D.V., Baryshnikov, A. Prognostic value of expression of antigen CD95 (Fas/APO-1) in patients with kidney transplantation // Medical immunology.- 2001,-v.3.- No. 2.-S-152.

6. Simonov, A.V., Arshinov S., Kozyreva O.V. Clinical value of determination of population composition of peripheral blood lymphocytes in various infectious processes // Medical immunology.-1999.-so 1.No. 3-4.-P.86.

7. The Yarylo A.A. fundamentals of immunology: a Textbook. - M.: Medicine, 1999.-608 C.

8. Bourn A. Isolation of mononuclear cells and granulocytes from human blood and bone marrow// Scand. J. Clin. Invest. - 1968. - V.2L - N97.- P.77.

A method for diagnosing an internal endometriosis in women by examining peripheral blood, characterized in that in the venous blood sample to determine the relative content of CD25+ lymphocytes and their values equal to 6% and more, diagnose internal endometriosis.



 

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