RU2308891C2 - Method for diagnosis of genitalia tuberculosis in women - Google Patents

Abstract

FIELD: medicine, phthisiology, gynecology.

SUBSTANCE: method involves sampling of biological material with maximal approaching to the tuberculosis inflammation focus. Method involves carrying out investigation of biological material by using chemiluminescence procedure wherein the level of luminol-dependent chemiluminescence of peritoneal liquid is determined before sampling segmentary tuberculin tests on anterior abdominal wall and in 72 h after carrying out. Tuberculosis is diagnosed in case of increasing the parent index by above 2 times. Proposed method provides enhanced reliability of diagnosis.

EFFECT: improved method of diagnosis.

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Description

The invention relates to medicine, namely to phthisiology, and can be used to diagnose genital tuberculosis in women.

The relevance of the modern diagnosis of female genital tuberculosis is determined by its significant place in the structure outside of pulmonary localization and the complexity of its detection in the early stages of the disease. Diagnostic difficulties are caused by small forms and the limited nature of the lesion, and later recognition leads to the development of significant anatomical changes, fibrosis and infertility. The primary lesion of the fallopian tubes significantly limits the collection of material, which significantly reduces the possibilities of examination methods. In this regard, the development of methods that allow quickly and reliably assess the presence of tuberculosis infection in the female genital organs and determine the degree of its activity is of particular relevance. Diagnostic statement of segmental samples (patent No. 2154415) implies an exacerbation of the tuberculous process on the introduction of a specific antigen, which is accompanied by local (skin) and focal (in the genitals) reactions. However, the focal reaction is not always perceptible, and local manifestations do not allow to localize the lesion. To objectify the samples, additional studies are needed: bacteriological, biochemical (ADA), immunological (ELISA) and morphological, which confirm the diagnosis according to the literature in 30%, and according to the data obtained by us over the past 5 years, in 10% of cases. With the isolated use of tests before and after tests in 100%, the diagnosis is confirmed only when Langhans cells are found in peritoneal fluid.

The prototype of the proposed method was a screening method for determining the CL of the cellular elements of menstrual blood (Gracheva M.P., Kraynenko E.V. Chemiluminescent method for the diagnosis of genital tuberculosis in women // Tuberculosis and Ecology. - 1997. - No. 3. - P.36-38 .). The test is carried out using a specific mitogen (BCG), in response to the addition of which in patients with tuberculosis, in contrast to patients with non-specific inflammatory processes, an increase in the level of blood chemiluminescence is noted. However, this method is not specific for tuberculosis of the female genital organs, since a recorded increase in chemiluminescence can be a reaction to any localization of the tuberculosis process, since the menstrual discharge includes peripheral blood with all its shaped elements.

The objective of the proposed method is to increase the reliability of the diagnosis of tuberculosis of female genitalia.

The task is carried out due to the fact that the activity of peritoneal fluid cells is studied using chemiluminescence before conducting segmental tuberculin tests on the anterior abdominal wall and after 72 hours, and when the initial indicator is more than doubled, tuberculosis is diagnosed.

The advantages of the method are the collection of biological material as close as possible to the focus of tuberculous inflammation, which ensures its high diagnostic accuracy. The method is specific for tuberculosis of female genitalia, since it is carried out against the background of segmental tuberculin samples, in which tuberculin is injected intradermally into the area of projection of the internal genitalia onto the anterior abdominal wall.

The method is as follows.

When staging the CL reaction, a 0.5 ml sample of the peritoneal fluid obtained by puncture of the Douglas space is placed in a sterile tube with RPMI 1640 medium (Biolot. SPb) with the addition of 20 IU of heparin per 1 ml. The study is carried out twice - before the tuberculin test and 72 hours after. Count the number of cells in punctate, in a light microscope in a Goryaev’s chamber. Luminescent chemiluminescence is recorded on a VIKTOR 2 1420 instrument (Wallas. Finland) in 96-well white opaque plates (Costar company). To determine the level of spontaneous chemiluminescence, 20 μl of punctate in medium, 40 μl of a solution of luminol 104 M, 140 μl of a balanced Hanks solution (Biolot) pH 7.2 are added to each well.

To place the induced sample, 20 μl of punctate in the medium, 20 μl of zymosan, opsonized with the blood serum of donors (or 20 μl of farbol myristate), 40 μl of luminol, 120 μl of Hanks solution are added to the well. The results are evaluated in dynamics over 40 minutes and expressed as light sum, i.e. pulses per second counted over the specified time.

This method allows you to objectively assess the development of the inflammatory process in the fallopian tubes in response to the introduction of tuberculin. A significant increase in chemiluminescence in response to the inducer after staging segmental samples in women with genital tuberculosis compared with patients with non-specific gynecological diseases was revealed (see table). A diagnostic criterion for an individual patient is considered to be an increase in induced CL by more than 2 times compared with the initial level.

Table Features of the chemiluminescent response of Duglasov punctate cells in female genital tuberculosis patients Patient groups Chemiluminescence induced by farbolmyristate (imp / s in 40 min) induced by zymosan (imp / s for 40 min) Before sampling After staging samples Before sampling After staging samples Non-specific diseases
N = 5 27.5 ± 6.7 38.6 ± 31.34 76.9 ± 27.2 73.5 ± 56.6 Tuberculosis
N = 8 8.6 ± 2 31.7 ± 10.3 ^* 38.9 ± 8.8 184.8 ± 70.9 ^*

^* - Differences in comparison with the corresponding indicators before the sampling.

The method has a high sensitivity of 87.5% and a specificity of 98.7%.

Clinical examples.

Example No. 1. Patient S., 32 years old, medical history No. 326; entered the State Institution "SPb NIIIF" in 2003. Complaints on admission to the impairment of menstruation since October 2002, infertility over the past 2 years, weakness, sweating, weight loss - 6 kg. Monthly from 13 years, regular, 5-6 days after 28-30, plentiful, painless. Since 2002, notes the impoverishment of menstruation. Objectively: The external genitalia are developed correctly. In the mirrors: the cervix and the vaginal mucosa of the usual color, the cervix with the phenomena of endocervicitis. Bimanual examination: the body of the uterus is not enlarged, mobile, painless. The right ovary is painless on palpation, not enlarged. To the left in the area of the appendages is weakly palpated, painful formation of irregular shape. The vault is shortened on the left.

Hysterosalpingogram: the uterine cavity is of normal size with uneven contours in the bottom area on the right. Pipes are contrasted on both sides for 3-4 cm, end with spherical extensions, impassable. In the cervical canal, uneven contours.

After staging segmental tests on the anterior abdominal wall, the patient developed pains in the lower abdomen, weakness, sweating intensified. A gynecological examination noted the appearance of pain on the left in the area of the appendages and the posterior fornix of the vagina, on the basis of which the focal reaction was regarded as positive. The general reaction is positive: after tuberculin tests, the patient has increased weakness, sweating. The local reaction to the introduction of tuberculin is negative. Serological reactions: PKK 47 srvc. units, RPG 33.0 srvc units, IFA 0.326 units. opt. tight - all are positive. IFA punctate Douglas space to samples of 0.426 units. opt. tight., after samples of 0.440 units. opt. tight, punctured ADA before samples 6.5 units, after samples 10.1 units, which are diagnostic criteria for tuberculosis. The level of winter-induced luminol-dependent chemiluminescence in a liquid from the Douglas space after setting up segmental samples was five times higher than the initial one, from 39.9 imp / s in 40 min to 205.4 imp / s in 40 min, the level of luminol-dependent chemiluminescence induced by farbol myristate increased three times from 10.0 pulses per second in 40 minutes to 34.7 pulses per second in 40 minutes, which is a diagnostic criterion.

Diagnosis: Tuberculosis of the uterus is active, the office is negative. Infertility II.

Example No. 2. Patient B. 27 years old, medical history No. 381; entered the clinic GU "SPb NIIIF". Complaints about the absence of pregnancy for 1.5 years. Monthly from 12 years to 6-7 days after 30, regular, painful on the 1st day, plentiful. In 2001, she began to be examined for the absence of pregnancy.

Objectively: The external genitalia are developed correctly. In the mirrors: Vaginal mucosa of the usual color, erosion on the cervix. Bimanual examination: the body of the uterus is enlarged, corresponds to 5/6 weeks of pregnancy, dense coarse-tuberous, mobile, sensitive to palpation. On the left, the appendages are not palpable, the paranasal region of their palpation is painful. An enlarged sensitive ovary is palpated to the right.

Hysterosalpingogram: the uterine cavity is slightly enlarged, deviates to the left, filling defects are not excluded in the area of uterine angles, mucosal hyperplasia, polyps. The left pipe is not contrasted. The right tube is thinned, contrasted to the ampullar region, turned into a sactosalpinx.

During the examination of the patient, a graded skin test and a graduated Mantoux test were delivered in V and VI dilutions on the forearms. The local reaction was within normergy. General and focal - negative. The patient received segmental tests on the anterior abdominal wall. The local reaction is negative. General and focal reactions are also not registered. The results of a comprehensive examination confirmed the non-specific nature of the pathological process. In this case, an increase in the level of chemiluminescence of peritoneal fluid was not registered.

Thus, the above examples illustrate the information content of the claimed method for the diagnosis of tuberculosis of the female genital organs.

Claims (1)

A method for the diagnosis of female genital tuberculosis, including the study of biological material using chemiluminescence, characterized in that the level of luminol-dependent chemiluminescence of peritoneal fluid is determined before conducting segmental tuberculin samples on the anterior abdominal wall and 72 hours after and when the initial indicator is more than 2 times diagnosed with tuberculosis.