RU2475752C1 - Method of predicting of pregnancy outcome in case of threat of miscarriage at late terms - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: in order to predict premature labour in pregnant women at 22-33 week term, relative content of CD62L+neutrophils is determined in peripheral blood and if its value is equal or is higher than 65%, premature labour is predicted.

EFFECT: application of the claimed method makes it possible to predict development of premature labour, which makes it possible to select correct tactics of managing women with risk of said obstetric pathology development.

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Description

The invention relates to medicine, namely to obstetrics, and can be used to predict premature birth in pregnant women with the threat of miscarriage at a gestational age of 22-33 weeks.

The relevance of the claimed method is determined by the fact that preterm birth is one of the main causes of perinatal morbidity and mortality. The frequency of this complication of pregnancy is 5-12% (Sidorova I.S., Makarov I.O. The course and management of pregnancy in trimesters. - M .: Medical Information Agency LLC, 2007. - P.304; Goldenberg R. et al. The Alabama preterm birth. Prevention project // Obstet. Gynecol. - 1990. - Vol. 75. - P.933-939). The development of this pathology leads to a high frequency of complications from the mother: abnormalities of labor (weakness, discoordination of labor), hypo- and atonic bleeding, injuries, infections; from the fetus - infectious complications, neurological disorders (Sidelnikova V.M. Preterm birth. Preterm baby. - M .: GOETAR-Media, 2006. - P.207).

Prediction of preterm labor will reduce the incidence of complications from the mother and fetus.

A known method for predicting the outcome of pregnancy in threatening preterm delivery, including a study of the peripheral blood of a pregnant woman from 28 weeks of gestation, in which before and after a course of preservation therapy quantitative indicator of nitrate ions is determined and with its increase by 10% or more relative to the initial value, the outcome of pregnancy is predicted in the form of preterm birth (patent No. 2272574. Russian Federation. A method for predicting the outcome of pregnancy with threatening preterm birth. / Possess va LV, Nazarov SB, SA Nazarov, Kuzmenko GN, Vorobyov YS // Inventions, Utility Models, 2006. -. №9).

The method has disadvantages:

- prediction of pregnancy outcome only from the third trimester;

- the need to examine the patient twice;

A known method for predicting preterm labor with urogenital infection, which includes the study of the cervical canal, is used to examine the epithelial cells of the mucous membrane, in which the expression level of the Toll-like receptor 2 (TLR2) gene is determined in real time by increasing the polymerase chain reaction more than five times in comparison with its level established during normal pregnancy, predict preterm delivery (patent No. 2334233. Russian Federation. Method prognostic preterm birth with urogenital infection. / Gankovskaya LV, Makarov OV, Kovalchuk LV, Bakhareva IV, Romanovskaya VV, Gankovskaya OA // Inventions. Useful models. - 2008. - No. 26).

The method has disadvantages:

- the study is associated with manipulations in the vagina and cervix, which can provoke untimely discharge of amniotic fluid and premature birth;

- the application of the method is limited to examination of women with urogenital infection (UMI);

- the need to determine laboratory norms for the level of TLR2 expression in the cervical canal of pregnant women;

- the non-specificity of the method, since an uncomplicated pregnancy can also occur against the background of an ultrasound scan, which in turn affects the expression of TLR2 in the cervical canal.

A known method for predicting the outcome of pregnancy in threatening preterm delivery by examining the peripheral blood of pregnant women with the threat of preterm delivery of the content of CD16 + and CD3 + cells. The ratio of their ratio is calculated by the formula CD16 + / CD3 +. When the values of indicators less than or equal to 0.11 predict the onset of preterm birth (patent No. 2132069. Russian Federation. A method for predicting the outcome of pregnancy with threatening preterm birth. / Kroshkina N.V., Sotnikova N.Yu., Bukina EA, Antsiferova Yu.S., Kudryashova A.V. // Inventions. - 1999. - No. 17, Part 2. - P.407).

This method was chosen by us as a prototype. However, it has several disadvantages:

- the method is time consuming, because requires the study of several types of cells and the calculation of the indicator by the formula;

- the accuracy of the method is 78%.

The technical result is to increase the accuracy of prediction of preterm birth in pregnant women in the period of 22-33 weeks with the threat of miscarriage.

The claimed technical result is achieved by the fact that in pregnant women with a risk of miscarriage in the period of 22-33 weeks of gestation, the relative content of CD62L + neutrophils in the peripheral blood is determined and at its value equal to or more than 65%, premature births are predicted with an accuracy of 90%, sensitivity of 90% and specificity of 90%.

The novelty of the proposed method lies in the fact that for the first time it is proposed to predict the outcome of pregnancy with the threat of miscarriage at a gestational age of 22-33 weeks by determining the relative content of CD62L + neutrophils in peripheral blood.

According to the literature, the role of CD62L + neutrophils has been little studied. It is known that the relative content of CD62L + neutrophils in peripheral blood increases after surgical procedures due to the regulatory role of pro-inflammatory cytokines (Ball C.J., King M.R., 2011). According to Kourtis A.P. et al. (2003) CD62L molecule is a marker of generalized neonatal infection. The role of CD62L + neutrophils in the initiation and development of allergic reactions is studied (Monteseirín J., Chacón P. et al., 2005). There is evidence of an increase in the relative content of CD62L + neutrophils in the amniotic fluid during intra-amniotic infection (Hsu C.D., Meaddough E. et al., 1998). Previously, the determination of CD62L + neutrophils in the peripheral blood of women with the risk of miscarriage in the period 22-33 weeks of pregnancy was not used to predict the development of preterm labor. There are no data in the literature on changes in the content of CD62L + neutrophils with the threat of miscarriage, depending on its outcome. It was established that neutrophils as the main producers of proteolytic molecules play a direct role in preparing the body for childbirth. It is known that CD62L molecules determine the interaction of cells with endotheliocytes, regulating the processes of cell exit from the vascular bed into the tissue (Laudanna C., 2006). Probably, an increase in the expression of CD62L molecules on the surface of peripheral blood neutrophils in women with the threat of miscarriage at 22-33 weeks of gestation is one of the mechanisms determining the enhanced migration of neutrophils into the cervical tissue immediately before unleashing labor.

The method is carried out according to the standard method as follows:

1. Add 3 ml of Medium 199 to 3 ml of heparinized blood from a cubital vein and isolate the enriched neutrophil population using the standard method of fast centrifugation in double density gradient ficoll-verographin (Bourn A. Isolation of mononuclear cells and granulocytes from human blood and bone marrow // Scand J. Clin. Invest. - 1968. - V.21. - N 97. - P.77).

2. A suspension of neutrophils is washed twice with saline, the cell concentration is adjusted to 1 × 10 ⁶ cells / ml. 5 μl of anti-human CD62L monoclonal antibodies conjugated to FITC (BD Bioscience Pharmingen, USA) are simultaneously added to 0.1 ml of the resulting suspension. Cells are incubated for 30 minutes at room temperature in the dark, washed in 1 ml of saline and fixed in 0.5 ml of CellFix ™ fixing solution (BD Biosciences, Belgium).

3. Determine the relative (percentage) content of CD62L + cells in a neutrophilic gate using a flow cytometer, and if it is equal to or more than 65%, preterm delivery is predicted.

Using the proposed method allows with high accuracy 90% to predict the outcome of pregnancy in women with the threat of miscarriage in the period of 22-33 weeks.

Distinctive features of the claimed method are:

- a new prognostic parameter was established for the relative content of CD62L + neutrophils in the period of 22-33 weeks of pregnancy, at a value of which 65% or more predict the development of preterm birth.

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

Example 1. Patient R., 25 years old, fourth pregnancy, desired, in a registered marriage. A history of 1 timely delivery without complications, 2 medical abortions without complications, and chronic pyelonephritis from extragenital pathology. The course of pregnancy was complicated by the threat of termination in the period of 22 weeks, about which she was in hospital treatment.

In the period of 22 weeks, a survey was conducted according to the claimed method:

The relative content of CD62L + neutrophils was 65%, which is equal to the claimed parameter (= 65%).

Conclusion: The development of preterm birth is predicted. Conservation therapy was carried out with effect. At the time of discharge, there were no signs of a threatened abortion.

In a period of 28 weeks, she was again hospitalized in the birth block. Diagnosis: Pregnancy 28 weeks. Head presentation of the fetus. The first period of preterm birth. Chronic pyelonephritis without exacerbation.

Preservation therapy has begun, with no effect.

A live premature boy was born weighing 930 g, height 34 cm, rating on the Apgar scale 4 points, on the Downs scale - 6 points, the newborn was transferred to the intensive care unit.

The forecast for the claimed method was confirmed.

Example 2. Patient N., 26 years old. Second pregnancy, desired. A history of 1 medical abortion, without complications. The course of pregnancy was complicated by the threat of termination at 12 weeks, inpatient treatment was carried out, with effect. In a period of 33 weeks about the threat of preterm birth, she was hospitalized in the department of pregnancy pathology.

A survey was conducted according to the claimed method:

The relative content of CD62L + neutrophils was 23%, which is lower than the claimed parameter (<65%).

Conclusion: Delivery is predicted on time.

Conservation therapy was carried out, with effect.

Pregnancy ended in timely delivery at 39-40 weeks of gestation. A live full-term girl was born, weighing 3350 g, height 51 cm, rating on the Apgar scale of 8-9 points.

The forecast for the claimed method was confirmed.

Example 3. Patient Z., 24 years old. Second pregnancy, desired. A history of 1 medical abortion, without complications, vegetovascular dystonia. The course of pregnancy was complicated by the threat of interruption in the period of 8 weeks, outpatient treatment was carried out with effect. In the period of 16 weeks suffered ARVI. In a period of 28 weeks, she was admitted to the Department of Pregnancy Pathology with the threat of premature birth.

A survey was conducted according to the claimed method:

The relative content of CD62L + neutrophils was 92%, which is higher than the claimed parameter (> 65%).

Conclusion: The development of preterm birth is predicted. Conservation therapy was carried out with effect. At the time of discharge, there were no signs of a threatened abortion.

At 36 weeks she was admitted to the obstetric clinic with regular labor. Diagnosed with Pregnancy 36 weeks. Head presentation of the fetus. 1 period of childbirth. Vegetovascular dystonia.

The first preterm birth occurred at 36 weeks.

A live premature girl was born weighing 2,400 g, growing 46 cm, estimated on the Apgar scale 7-8 points, on the Downs scale - 2 points. From the maternity hospital, the newborn was transferred to the second stage of nursing of premature babies.

The forecast for the claimed method was confirmed.

The research results are summarized in table.

Indicator The number of examined Total women thirty True positive result 9 False positive 2 True negative result eighteen False negative result one Total: the accuracy of the proposed method is 90% The sensitivity of the proposed method is 90% The specificity of the proposed method is 90%

The advantages of the proposed method:

1. High accuracy - 90%;

2. Good reproducibility of the method;

3. Simplicity in interpretation of survey results;

4. The method allows to predict preterm birth at certain periods of pregnancy (22-33 weeks), which makes it possible to prevent this complication of pregnancy and to prevent the occurrence of both complications of labor and perinatal pathology.

Claims (1)

A method for predicting the outcome of pregnancy with the threat of miscarriage by determining immunocompetent cells in the peripheral venous blood, characterized in that the relative content of CD62L + neutrophils is determined in the gestational age of 22-33 weeks, and if it is equal to or more than 65%, preterm delivery is predicted.