Early diagnosis method for diagnosing external genital endometriosis in women

IPC classes for russian patent Early diagnosis method for diagnosing external genital endometriosis in women (RU 2247391):

G01N33/68 - involving proteins, peptides or amino acids

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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.

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The invention relates to medicine, namely to gynecology, and allows you to diagnose external endometriosis at the preclinical stage.

The urgency of the invention is determined by a high frequency of occurrence (50%) of external endometriosis in the structure of gynecological morbidity [1, 2], its Association with impaired reproductive function [1, 5], and thus the influence on the health of not only women, but also the family.

Currently, the diagnosis of external endometriosis is established histologically by biopsy at the time of surgical intervention during laparoscopy or laparotomy.

A known method for the diagnosis of genital endometriosis by examining the functional activity of basophils in the peripheral blood of the patient in the presence of an aqueous extract of endometrial tissue protein concentration >200 μg/ml With >12% of diagnosed genital endometriosis [4].

The disadvantages of the method:

- allocation of basophilic leukocytes requires sampling a large number of blood (18 ml) due to the relative small number of basophils in the peripheral blood (0.5 to 1%);

- proposed to use the extract endometriotic tissue may not be standardized and requires testing after each extraction and are not included in the Register of medical products and allowed to skin the of MOH”;

the method does not allow for diagnosis of various forms of genital endometriosis (the method used for the diagnosis of both early and clinically significant forms);

- do not specify the accuracy of the method.

Closest to the claimed method is a method of diagnosis of endometriosis by detecting the endometrial biopsy taken at 20-22 days of the menstrual cycle, activity gidroksietilirovanny and lactate dehydrogenase, calculating their ratio. The diagnosis of endometriosis set at the value less than 0.7 [As the USSR, ICB G 01 N 33/48. The way to diagnose endometriosis / Vasoli, Tagetrov, Tashenov (USSR). No. 3738785 (28-14) // Discoveries and inventions. -1986. No. 44. -s].

This method elected us as the nearest equivalent. However, this method has the following disadvantages:

- How complex in execution due to the need to study two biochemical parameters in the endometrium.

- Complicated the interpretation of these data because the end result is obtained by mathematical calculations and not the simple reading of the indicator device.

- Not known the possibility of using this method to diagnose early, preclinical forms of external endometriosis.

- Do not specify the accuracy of the method.

The disadvantages of this method of diagnosis is proposed device is a thread in the claimed method.

The technical result of the proposed method lies in the fact that women in the biopsy of the endometrium investigated the activity of cathepsin D and the values from 0.1 Adherent/hour and less external endometriosis is diagnosed with an accuracy of 82.6%.

The method is standard [3] as follows: bar gets endometrial biopsy (ZUG), homogenized in a laboratory mortar, then the sample of endometrial tissue 0.1 g suspendered in 0.25 ml of acetate and 2.8 H buffer in two test tubes. Then in each tube is made of 0.25 ml of 1% solution of hemoglobin. In one of the tubes immediately paid 5 ml of 10% solution of trichloroacetic acid (control). Both tubes are incubated at t°=+37°C for 60 minutes. After a specified time in the second test tube also added 5 ml of trichloroacetic acid and both tubes centrifugeuse for 30 minutes at 3000 g. Then determined the optical density of the obtained supernatant on a spectrophotometer SF-46” at a wavelength of 280 nm against control. The result is expressed in units of optical density, where every 0.1 eop correspond to 0.1 Adherent./hour.

Previously, the determination of the activity of cathepsin D in the blood and tissues was used as an indicator of invasive potential of malignant tumors and is useful in the diagnosis of gastric cancer and breast dakoda this time its clinical efficacy is not defined [6].

Novelty: for the first time cathepsin D used for preclinical diagnosis of early forms of external endometriosis.

Distinctive features of the method are:

Established diagnostic activity of cathepsin D to external endometriosis, the endometrial biopsy in the range from 0.1 Adherent/hour or less.

The essence of the method is illustrated by the following examples:

1. Patient B-VA, 27 years. Entered the survey in SE "Ivanovo research Institute of the Ministry of Foreign Affairs to them. Vnegorodov" health Ministry with a diagnosis of Primary infertility. During examination, the patient was excluded immunological, endocrine, inflammatory and male factors of infertility in tests of functional diagnostics, the level of prolactin, hysterosalpingography, positive postcoital test, the sperm of the husband. When taking Zug endometrium it was determined the activity of cathepsin D by the present method, which was 0,058 Adherent/hour, which is less than the proposed option (0,1 Adherent./h) in the presence of external genital endometriosis.

Given the lack of data for the apparent cause of infertility, the patient was recommended and subsequently performed a diagnostic laparoscopy, where he was diagnosed with external endometriosis stage I (on the pelvic peritoneum were found sporadic endometrioid heterotopia, the total area of pathologic what about the process up to 3 cm ²), verified according to the endometrial biopsy heterotopia.

Conclusion: the diagnosis of external genital endometriosis claimed method was confirmed.

2. Patient G-d, 35 years. Entered the gynecological clinic of SE "Ivanovo research Institute of the Ministry of Foreign Affairs to them. Vnegorodov" health Ministry with a diagnosis of MFD by type algodonera, suspected internal endometriosis.

The patient was performed hysteroscopy, in which the internal endometriosis was removed. Simultaneously biopsy of the endometrium, where he determined the activity of cathepsin D, which amounted to 0.10 Adherent/h, which corresponds to the upper boundary of the proposed option (0,1 Adherent/h) external genital endometriosis.

The patient in the next menstrual cycle was performed diagnostic laparoscopy, the indication for which was persistent pain resistant to treatment. On operations was diagnosed with external endometriosis stage II (on the Sacro-uterine ligaments and Dupacova space were found deep foci of endometriosis, with a total area of more than 3 cm²), verified according to the endometrial biopsy heterotopia.

Conclusion: the diagnosis of external genital endometriosis claimed method was confirmed.

3. Patient G-VA, 32 years. Entered in the state, "Ivanovsky research Institute of the Ministry of Foreign Affairs to them. Vnegorodov" health Ministry with diag is oz: secondary infertility. During examination, the patient was excluded: immunological, endocrine and male factors of infertility in tests of functional diagnostics, the level of prolactin, Zug endometrium, positive postcoital test, the sperm of the husband. When taking Zug endometrium it was determined the activity of cathepsin D by the present method, which amounted to is 0.102 Adherent/h, which does not match the declared parameter (0,1 Adherent/h) external genital endometriosis. Hysterosalpingography was an infringement of the output of the contrast agent from the lumen of the fallopian tubes. Was suspected chronic salpingitis, about which the patient was performed diagnostic laparoscopy. The diagnosis of chronic salpingitis was confirmed: the fallopian tube had sharply pronounced crimp, emerging phimosis fibrillose Department. Abnormalities in the pelvic peritoneum and the abdominal cavity was not found.

Conclusion: the absence of a diagnosis of external genital endometriosis claimed method was confirmed, although infertility has been caused by another factor - chronic salpingitis.

In this way were examined 92 women, one of them positive results by the present method were obtained in 76 people, that is to 82.6%. Diagnosis of external endometriosis verified when the paw is octopii and biopsy of endometrioid heterotopy. The results of the study are shown in table 1.

The advantages of the proposed method:

1. The ability to diagnose preclinical stage endometrioid process.

2. Automated reading of research results, eliminating subjectivity in the evaluation of the result.

3. Simplicity and low cost method.

Table 1. Confirmation of the diagnosis by the present method.
All surveyed	A positive result	The accuracy of the method
92 women	76 people	' p<0,01

References

1. L.V. Adamyan, fists Century. And. Endometriosis: a Guide for physicians. -M.: Medicine. -1998. -320 C.

2. State report on the state of health of the Russian Federation in 2000, / / “the public Health of the Russian Federation”. -2002. No. 3. -p.8.

3. Complementary mechanism: research methods / edited by j. Dingle // M.: Mir. -1980. -344 S.: ill.

4. Patent No. 2004910. Kulakov V.I., Sukhikh G.T., Pshenichnikova TA, Volkov, N.I., Ions I.D. of the Russian research center of Perinatology, obstetrics and gynecology. Method for the diagnosis of genital endometriosis. // The Bulletin “Inventions”. -1993. No. 45-46. -164.

5. The summary A.N., Davydov, A.I. Endometriosis. CL the technical and theoretical aspects. // M: Medicine. -1996. -330 C.

6. Encyclopedia of clinical laboratory tests / Ed. by Tietz N. // TRANS. from English. edited Riv. -M.: Publishing house "Labelform". -1997.-960 C.

The method of early diagnosis of external genital endometriosis in women by exploring endometrial biopsy, characterized in that the tissue sample to determine the activity of cathepsin D, and when it is 0.1 Adherent/h and less diagnose external genital endometriosis.