RU2587781C1 - Method of diagnosing severe preeclampsia - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention can be used for diagnosing severe preeclampsia in pregnant women. Method involves the peripheral venous immunoassay during 22-37 weeks of pregnancy. There is determined a relative content of CD62L + cells in the population of neutrophils and if its value is equal to or less than 57 %, severe preeclampsia is diagnosed.

EFFECT: use of the invention enables diagnosing of preeclampsia regardless of presence in a woman of chronic arterial hypertension.

1 cl, 1 tbl, 3 ex

Description

The invention relates to medicine, namely to obstetrics, and can be used to diagnose severe preeclampsia in pregnant women during gestation periods of 22-37 weeks.

The urgency of the problem is caused by the need to search for additional criteria for assessing the severity of preeclampsia in order to develop adequate management tactics for pregnant women and reduce the frequency of adverse pregnancy outcomes with this complication. The difficulty in assessing the severity of preeclampsia can be determined by several reasons. In a number of cases, against the background of initially high blood pressure in women with chronic arterial hypertension, it is not always clinically possible to differentiate the severity of joining preeclampsia. In addition, in clinical practice there are many cases of the erased, low-symptomatic course of preeclampsia, when the true severity of preeclampsia cannot be assessed by the general condition, blood pressure and proteinuria. Thus, the imperfection of objective methods for the diagnosis of preeclampsia leads to an underestimation of the true severity, later diagnosis and, as a consequence, the ineffectiveness of therapeutic measures.

Therefore, one of the main tasks in determining the management of pregnant women with preeclampsia is the identification of additional diagnostic criteria for the severity of this pregnancy complication.

Currently, there are methods to diagnose preeclampsia and determine the severity. However, most of them do not take into account the presence of chronic arterial hypertension in a pregnant woman.

There is a method for diagnosing severe gestosis, characterized in that they monitor blood pressure and heart rate during the day (RU 2002105984 A, Guryev V.M. et al., 20.11.2003). According to the results of the study, each of the measured diastolic pressures is correlated with the maximum allowable for a given heart rate (HR), calculated by the formula ADdiast = [- 0.569 + 17.7 * log (heart rate)]. In the case when the excess of the maximum allowable diastolic pressure is observed in more than 55% of measurements during the day and / or more than 70% at night, severe gestosis is diagnosed.

The disadvantages of the method: the method is time-consuming, the duration of the diagnostic process delays the correct diagnosis, there is no possibility of diagnosing preeclampsia in women with chronic arterial hypertension due to the possibility of obtaining a false-positive result.

There is a method for the diagnosis of gestosis by determining the level of alpha-amylase in the blood serum by a quantitative method (RU 2426129 C1, Grebenkin B.E. et al., 10.08.2011). At an alpha-amylase level of 28-100 U / L, mild gestosis is diagnosed. At an alpha-amylase level> 100 Units / L, severe gestosis is established. The disadvantages of the method include the possibility of obtaining high values of alpha-amylase and, therefore, a false-positive result in the presence of diseases of the pancreas. In addition, there is no data on the accuracy, specificity and sensitivity of this method, as well as the possibility of diagnosing preeclampsia by this method in women with chronic arterial hypertension.

A known method for the diagnosis of severe forms of gestosis by determining the level of homocysteine in plasma of venous blood (RU 2259568 C2, Sukhikh G.T. et al., 27.08.2005). With an increase in homocysteine levels more than 2.2 times the norm, the development of severe gestosis is diagnosed. The disadvantage of this method is that homocysteine is not a specific marker of preeclampsia, its level can increase in various conditions. Thus, insufficient intake of folic acid and B vitamins from the outside, which are enzymes necessary for the metabolism of homocysteine, can increase its level. In addition, hyperhomocysteinemia is a risk factor for the development of atherosclerosis and cardiovascular diseases (Frank, J. No evidence for prooxidative effects of homocysteine in vascular endothelial cells / J. Frank, SC Beck, A. Flaccus, HK Biesalski // Eur J Nutr. - 2007. - Vol. 46, No. 5. - P. 286-292; Sidorenko G.I., Moisinok A.G. The role of homocysteine in thrombogenesis and atherogenesis // Cardiology, Abstract: doc. - M., 2002 - P. 56-61), which does not allow the use of this method for the diagnosis of preeclampsia in women with chronic hypertension.

There is a method for diagnosing severe gestosis based on determining the concentration of the hormones leptin and ghrelin in venous blood by enzyme immunoassay (RU 2476880 C1, Shkurat T.P. et al., 02.27.2013). With an increase in the concentration of the hormone leptin in the blood serum of more than 37.89 ng / ml and a simultaneous decrease in the concentration of the hormone ghrelin in the blood plasma below 0.26 ng / ml, severe gestosis is diagnosed.

The disadvantages of the method is that an increase in the level of leptin can be associated with the presence of obesity, metabolic syndrome, diabetes mellitus and essential hypertension (Tender features of the concentration of leptin and intrarenal hemodynamics in patients with essential arterial hypertension and obesity. / V.I. Podzolkov, A. E. Bragin, Yu.N. Rodionova // Systemic hypertension. - 2011. - No. 2, P. 42-46). This may contribute to the overdiagnosis of preeclampsia and the inability to diagnose preeclampsia in women with chronic hypertension. This method has diagnostic value only if two indicators change simultaneously. The accuracy of the method is 78.3%. In addition, this method of diagnosis is possible only in the third trimester of pregnancy, while the development of preeclampsia is possible in the earlier terms, even after 20 weeks of pregnancy.

Closest to the technical solution to the claimed method is the "Method for assessing the severity of preeclampsia in pregnant women" (RU 2250468 C2, Levchenko V.G. et al., 20.04.2005). This method is selected as a prototype. The essence of the invention lies in the fact that in the blood serum of pregnant women determine the content of the circulating immune complex of the main protein of myelin with class G immunoglobulin during pregnancy after 20 weeks and its values less than 0.1 μg / ml indicate an uncomplicated pregnancy, a level of 0.1 up to 0.2 μg / ml - about mild preeclampsia, a level of 0.2 to 0.5 μg / ml - about severe preeclampsia. However, a different classification of hypertensive disorders during pregnancy is currently used in clinical practice, according to which moderate preeclampsia and severe preeclampsia are distinguished (Edema, proteinuria and hypertensive disorders during pregnancy, childbirth and the postpartum period (O10-O16) // International statistical classification of diseases and health problems. Tenth revision). Accordingly, this method does not take into account modern diagnostic criteria, on the basis of which the severity of preeclampsia is determined. In addition, the description of this method does not provide information on the possibility of diagnosing preeclampsia in women with chronic hypertension.

The technical result of the patented method are: expanding the arsenal of diagnostic tools for severe preeclampsia, compliance of the patented method for diagnosing the modern classification of hypertensive disorders during pregnancy, the possibility of diagnosing preeclampsia, regardless of whether a woman has chronic arterial hypertension.

The claimed technical result is achieved by the fact that in pregnant women during gestational periods of 22-37 weeks in the peripheral venous blood, an indicator of the relative content of CD62L + cells in the neutrophil population is determined and, with its value equal to or less than 57%, severe preeclampsia is diagnosed.

The novelty of the proposed method lies in the fact that for the first time it is proposed to diagnose severe preeclampsia in pregnant women, regardless of the presence of chronic arterial hypertension, as determined in the peripheral venous blood of a pregnant woman, the relative content of CD62L + cells in the neutrophil population.

There is evidence in the literature that a change in the content of soluble forms of CD62L molecules in women in early pregnancy is further associated with the development of preeclampsia (Adhesion molecules changes at 20 gestation weeks in pregnancies complicated by preeclampsia / Chavarria ME [et al.] / / Obstet Gynecol Reprod Biol. - 2008; 137 (2): 157-64).

A decrease in the level of CD62L + neutrophils in women with severe preeclampsia is possible as a result of elution (shedding) of L-selectin molecules under the influence of inflammatory mediators (Meager A. Cytokine regulation of cellular adgesion molecule expression in inflammation / Meager A. // Cytokine Growth Factor Rev. 1999; 10; 27-39). This can inhibit cell migration into tissue and promote the circulation of neutrophils activated during preeclampsia in the peripheral bloodstream (Interactions between inflammatory and oxidative stress in preeclampsia / Ouyang YQ [et al.] // Hypertens Pregnancy. 2009, Vol. 28, No. 1, Pages 56-62). Activated neutrophils produce a high level of reactive oxygen species that can block the vasodilatory effects of nitric oxide, thereby contributing to severe vasospasm in severe preeclampsia.

The method is as follows:

1) In a pregnant woman, venous blood from the cubital vein is sampled once, 100 μl of heparinized venous blood is incubated with 5 μl of monoclonal anti-CD62L labeled with FITC for 30 minutes at room temperature in the dark, after which red blood cells are lysed with standard solutions of manufacturers ( OptiLyse B, Beckman Coulter; FACS Lysing Solution, BD Biosciences et al.). Cells are washed 1 time with 1 ml of physiological saline by centrifugation at 1500 rpm for 2 minutes and resuspended in 500 μl of physiological saline. The reaction is recorded on a flow cytometer, determining the relative content of CD62L + cells in a neutrophilic gate. The result is read in percent (%). If the relative content of CD62L + cells in the neutrophil population is equal to or less than 57%, severe preeclampsia is diagnosed. The time of the study is 1 hour.

Using the proposed method allows with high accuracy - 85.2%, sensitivity - 81.5%, specificity - 88.9% to determine severe preeclampsia.

Distinctive features of the method are:

a boundary criterion for the relative content of CD62L + cells in the neutrophil population was revealed, at a value of which equal to or less than 57%, severe preeclampsia is diagnosed.

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

Example 1. Woman R., 42 years old, 33 weeks pregnant at the doctor’s appointment in the antenatal clinic. In somatic history - the presence of chronic arterial hypertension over the past 2 years. The initial blood pressure when registering is 140/100 mm Hg. Art., receives antihypertensive therapy (dopegitis). On examination - swelling of the legs and hands; blood pressure 160/100 mm Hg; in the general analysis of urine - proteinuria 0.33 g / l. A woman was sent to a hospital with a diagnosis of Pregnancy 33 weeks. Chronic hypertension with associated proteinuria. When examining the claimed method, it was found that the CD62L + cell count in the neutrophil population was 50%, and severe preeclampsia was diagnosed. In the hospital, a combination antihypertensive therapy, magnesia therapy was prescribed. On the background of treatment - blood pressure in the range 130 / 80-150 / 100 mm Hg, daily proteinuria 3 g / l, decreased urine output. Ultrasound examination shows signs of placental insufficiency; according to cardiotocography - a decrease in the compensatory abilities of the fetus. Given the presence of severe preeclampsia, the absence of the effect of ongoing therapy and intrauterine suffering of the fetus, it was decided to prepare the pregnant woman for early delivery.

Example 2. Woman N., 30 years old, pregnancy 22 weeks. Delivered by ambulance to an obstetric clinic with complaints of headache and high blood pressure up to 160/100 mm Hg. after psycho-emotional stress. The presence of chronic somatic pathology denies. During pregnancy, blood pressure is in the range 110 / 70-130 / 80 mm Hg. Upon receipt, blood pressure 160/110 mm Hg, urine - proteinuria 0.03 g / L. She was hospitalized in the intensive care unit with a diagnosis of severe preeclampsia. When examining the claimed method of the indicator of the content of CD62L + neutrophils, it was found that its level was 62%, severe preeclampsia was not detected. The pregnant woman was prescribed antihypertensive therapy (nifedipine, dopegitis). During the observation, blood pressure stabilized at values of 120 / 70-130 / 90 mm Hg, proteinuria was not detected in dynamics, the condition of the fetus was satisfactory according to an additional examination. Pregnancy was prolonged, the woman received antihypertensive therapy, was under dynamic observation in a stable satisfactory condition. Delivered on time.

Example 3. Woman A., 28 years old, pregnancy 37 weeks. I entered the department of pregnancy pathology of the obstetric clinic with complaints of high blood pressure. In somatic history - the presence of chronic pyelonephritis, obesity of 1 degree. During pregnancy, blood pressure within 120 / 70-130 / 80 mm Hg Upon enrolment

- blood pressure 150/100 mm Hg, in a single portion of urine

- proteinuria 1.16 g / l. Diagnosed with moderate preeclampsia. The woman is prescribed antihypertensive and magnesia therapy. When examining the claimed method of the indicator of the content of CD62L + neutrophils, it was found that its level was 57%, diagnosed with severe preeclampsia. During treatment, blood pressure in the range 140 / 90-150 / 100, proteinuria in the daily amount of urine - 5 g / l. According to an additional examination - a decrease in the compensatory capabilities of the fetus. It was decided to prepare the pregnant woman for delivery.

In this way, 54 pregnant women with preeclampsia were examined, of which 20 women with preeclampsia against the background of chronic hypertension. In 25 of the examined women, the content of CD62L + neutrophils was equal to or less than 57%, which corresponded to the diagnostic criterion for severe preeclampsia. Moreover, a false-positive result was obtained in 3 cases out of 25. 29 women were not diagnosed with severe preeclampsia, since the content of CD62L + neutrophils was more than 57%; in 5 women out of 29, a false negative result was obtained during the examination.

The accuracy of diagnosis of severe preeclampsia was 85.2%, sensitivity - 81.5%, specificity - 88.9%. The research data are shown in table 1.

The advantages of the proposed method:

1. the ability to diagnose severe preeclampsia in women with chronic arterial hypertension;

2. high accuracy, sensitivity and specificity;

3. a small time interval for the study;

4. simplicity and accessibility of the implementation of the methodology;

5. this method takes into account modern diagnostic criteria, on the basis of which the severity of preeclampsia is determined.

Claims (1)

A method for the diagnosis of severe preeclampsia in pregnant women, including an immunological study of peripheral venous blood at 22-37 weeks of pregnancy, characterized in that the relative content of CD62L + cells in the neutrophil population is determined and, with a value of equal to or less than 57%, severe preeclampsia is diagnosed.