RU2530586C1 - Method for prediction of anaemia in pregnant women - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: sampled peripheral blood is examined for an erythrocyte count containing no more than 5 formazan granules in one erythrocyte according to the cytochemical determination of glucoso-6-phosphate dehydrogenase activity. If observing an increase of the count up to 25-40%, developing anaemia is predicted. The presented method is easy to reproduce, requires no material inputs, enables predicting developing anaemia, reduces the time of laboratory findings and can be implemented in health care facilities.

EFFECT: method enables predicting anaemia in pregnant women in 90 per cent of cases.

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Description

The invention relates to medicine, namely to obstetrics, and can be used to predict, the development of anemia in pregnant women.

Among extragenital diseases complicating the course of pregnancy and childbirth, anemia occupies a leading place. About 90% of all anemia in pregnant women is iron deficiency anemia (IDA) [Shekhtman, M.M. Iron deficiency anemia and pregnancy // Gynecology. - 2006. - No. 6. - S.164-170]. Anemia leads to placental insufficiency, fetal growth retardation, as well as to a high percentage of premature births [Sidorova IS, Unanyan AL Miscarriage: violation of antioxidant defense and its correction // Russian Bulletin of the Obstetrician-Gynecologist. - 2009. - No. 1. - S.14-16].

A known method for the diagnosis of IDA by determining the concentration of ferritin in serum by enzyme immunoassay [Lee M., Burgett M.W. A solid phase enzyme immunoassay for the quantitation of serum ferritin // Clin. Chim. Acta. - 1981. - Vol. 112 (2). - P.241-246]. At a serum ferritin concentration of less than 15 μg / L, an iron deficiency condition is diagnosed. The disadvantages of this method are: nonspecificity (an increase in the concentration of serum ferritin in any infectious and inflammatory processes); in late pregnancy, the method is not informative; high cost of the method; due to the high cost and complexity of diagnosis, the proposed method is not used in the clinical practice of antenatal clinics and maternity hospitals.

A known method for the diagnosis of latent iron deficiency by determining the concentration of transferrin receptors in the blood plasma [Carriaga M. Serum transferrin receptor for detection of iron deficiency in pregnancy // Am. J. Clin. Nutr. - 1991 .-- Vol. 54. - P.1077-1081]. An increase of more than 13.5 mg / L is an early sign of iron deficiency during pregnancy. The disadvantage of the proposed method is its low specificity. The level of transferrin receptors can increase with an increase in the formation of red blood cells, which reduces the specificity of this method.

A known method for the diagnosis of iron deficiency by studying the morphology of red blood cells in fixed smears of peripheral blood [Grebennikova LG Diagnosis of the early stage of iron deficiency // InVestRegion. - 2008. - No. 1. - S. 57-59]. This method is chosen by us as a prototype. The method has significant disadvantages: the lack of reliable parameters for measuring the shape of red blood cells; the impossibility of use in clinical practice due to subjectivity in determining the morphology of red blood cells; Diagnostic results depend on the quality of preparation.

The task of the invention is to develop a technically simpler and more accurate method for predicting anemia in pregnant women.

The technical result is a simple, low-cost method based on the identification of ineffective erythropoiesis as a result of a decrease in the activity of glucose-6-phosphate dehydrogenase (G-6-FDG).

The technical result of our proposed method is achieved based on the identification of ineffective erythropoiesis and the formation of low-activity red blood cells. G-6-FDG is the main enzyme of the pentose phosphate pathway for glucose conversion, the only source of the reduced form of nicotinamide adenine dinucleotide phosphate (NADP) for adequate antioxidant protection of red blood cells. With a deficiency of G-6-FDG in red blood cells, the ability to convert oxidized glutathione to reduced is impaired, as a result of which the glutathione peroxidase antioxidant activity of red blood cells is suppressed. As a result of this, red blood cells are easily hemolysis under the influence of oxidizing agents with the development of anemia [Kozar R.A., Weibel C.J., Cipolla J. et al. Antioxidant enzymes are induced during recovery from acute lung injuri. - Critical Care Medicine. - 2000. - No. 28 (7). - P.86-91]. The decrease in the activity of G-6-FDG in red blood cells, determined by the intensity of the formation of granules of formazine in red blood cells, allows us to assess the degree of ineffective erythropoiesis and, based on this, predict the development of anemia.

The technical result is achieved in that according to the invention, when the number of red blood cells containing no more than 5 granules of formazan in one red blood cell is increased to 25-40% by the cytochemical determination of glucose-6-phosphate dehydrogenase activity, the development of anemia is predicted.

The proposed method is as follows.

Blood collection tubes are preheated at 37 ° C. 0.05 μl was taken from the examined patients from the finger, transferred into test tubes and mixed with 0.1 μl of a buffer solution consisting of glucose-6-phosphate disodium salt - 40 mg, NADP - 15 mg, MgCl ₂ - 200 mg , n-nitrotetrazolium violet 20 mg, methylene blue 10 mg, Richter heparin 25000 IU - 0.5 mg in 10 ml and Tris buffer with pH = 8.0. Blood in the buffer solution is kept in an thermostat at 37 ° C for 30 minutes. Then a blood smear is prepared, dried and microscopic, using an eyepiece with a magnification of 10 and a lens of × 90. Microscopy of smears counts the number of granules of formazan in each of 100 red blood cells. Determine the percentage of low activity red blood cells that contain 5 or less granules of formazan. At 25-40% of a low-activity population of red blood cells, anemia in pregnant women is predicted.

The low number of granules of formazan in red blood cells indicates an ineffective erythropoiesis, a breakdown of adaptive mechanisms in the body, which allows predicting the development of anemia in patients.

Examples of practical use of the proposed method.

Example 1

Pregnant V., 25 years old (individual pregnancy card No. 683, birth history 213), was registered at the antenatal clinic for 8 weeks. From the anamnesis it is known that menstrual function in a woman from 13 years old was established immediately. Menses 4 days, after 29 days, regular, painless. Last menstruation 05/14/2010. Gynecologically healthy. Sexual life since 18 years. In a registered marriage. Real pregnancy is the second. The previous pregnancy ended with an urgent birth in 2010, a boy was born weighing 3450, height 50 cm. Pregnancy proceeded against the background of anemia, childbirth was complicated by the early outflow of amniotic fluid, the postpartum period was complicated by hypogalactia.

Based on the anamnesis, a woman is at risk of developing anemia. A woman was studied the activity of G-6-FDG. Blood collection tubes were preheated at 37 ° C. The patient took blood from the finger in an amount of 0.05 μl, transferred to a test tube and mixed with 0.1 μl of a buffer solution consisting of glucose-6-phosphate disodium salt - 40 mg, NADP - 15 mg, MgCl ₂ - 200 mg, violet n-nitrotetrazolium 20 mg, 10 mg methylene blue, Richter heparin 25,000 m. - 0.5 mg in 10 ml and 0.02 μl of Tris buffer with pH = 8.0. Blood in a buffer solution was kept in an thermostat at 37 ° C for 30 minutes. Then a blood smear was prepared, dried and microscopic, using an eyepiece with a magnification of 10 and a lens of × 90. Microscopy of the smear counted the number of granules of formazan in each of 100 red blood cells. The percentage of low-activity erythrocytes containing 5 or less formazan granules was determined. The population of low activity red blood cells was 33%. The proposed method allowed to predict the development of anemia.

After 3 weeks, a laboratory study showed: OAK (erythrocytes 3.0 × 1012 / l, HB - 105 g / l, serum iron 11.3 μmol / l, the population of low-activity red blood cells - 38%. Anemia was registered in the patient.

Early delivery (36 weeks) was complicated by secondary weakness of labor, about which oxytocin was reinforced. It was delivered by a live premature baby weighing 2600 g, height 45 cm, with an Apgar score of 7/7 points. The postpartum period was complicated by hypogalactia.

Example 2

Pregnant V., 30 years old (individual pregnant card number 1023, birth history 108), was registered in the antenatal clinic for a period of 9 weeks. From the anamnesis it is known that menstrual function in a woman from 15 years old was established immediately. Menstruation for 5 days, after 30 days, regular, painless. Gynecologically healthy. Sexual life since 18 years. In a registered marriage. Real pregnancy is the first, desired. Profession - seller. The environmental background of the residence is unfavorable. Of extragenital diseases, chronic pyelonephritis is noted.

Based on the anamnesis, a woman is at risk of developing anemia.

A woman was studied the activity of G-6-FDH in red blood cells. Blood collection tubes were preheated at 37 ° C. The patient took blood from a finger in an amount of 0.05 μl and mixed with 0.1 μl of a buffer solution consisting of glucose-6-phosphate disodium salt - 40 mg, NADP - 15 mg, MgCl ₂ - 200 mg, n-nitrotetrazolium violet 20 mg, methylene blue 10 mg, Richter heparin 25,000 m. - 0.5 mg in 10 ml and 0.02 μl of Tris buffer with pH = 8.0. Blood in a buffer solution was kept in an incubator at 37 ° C for 30 minutes. Then a blood smear was prepared, dried and microscopic using an eyepiece with a magnification of × 10 and a lens of × 90. Microscopy of the smear counted the number of granules of formazan in each of 100 red blood cells. The percentage of low-activity erythrocytes containing 5 or less formazan granules was determined. The population of low activity red blood cells was 14%. The prognosis of pregnancy is favorable, without the development of anemia.

Childbirth ended on time, without complications. A live full-term baby was born weighing 3,500 g, height 52 cm, with an Apgar score of 8/8 points.

Example 3. Pregnant S., 33 years old (individual pregnancy card No. 1083, birth history 413), registered at the maternity welfare clinic for a period of 6 weeks. From the anamnesis it is known that menstrual function in a woman from 14 years old was established immediately, menstruation for 5 days, after 30 days, regular, profuse, painless. Gynecological diseases - uterine myoma. In a registered marriage. Real pregnancy is the second, desired. The previous pregnancy ended with an urgent birth in 2005, a boy was born weighing 3,750, 52 cm tall. The pregnancy was accompanied by anemia, the birth was complicated by discoordination of labor, the postpartum period was complicated by early hypotonic bleeding.

Based on the anamnesis, a woman is at risk of developing anemia. A laboratory study was carried out: red blood cells 3.5 × 1012 / l, HB - 123 g / l, serum iron 18.2 μmol / l.

A woman was studied the activity of G-6-FDH in red blood cells. Blood collection tubes were preheated at 37 ° C. The patient took blood from the finger in an amount of 0.05 μl and mixed with 0.1 μl of a buffer solution consisting of glucose-6-phosphate disodium salt - 40 mg, NADO - 15 mg, MgCl ₂ - 200 mg, n-nitrotetrazolium violet 20 mg, methylene blue 10 mg, Richter heparin 25,000 m. - 0.5 mg in 10 ml and 0.02 μl of Tris buffer with pH = 8.0. Blood in a buffer solution was kept in an incubator at 37 ° C for 30 minutes. Then a blood smear was prepared, dried and microscopic using an eyepiece with a magnification of × 10 and a lens of × 90. Microscopy of the smear counted the number of granules of formazan in each of 100 red blood cells. The percentage of low-activity erythrocytes containing 5 or less formazan granules was determined. The population of low activity red blood cells was 36%. The prognosis is poor, anemia may develop. The obtained indicators indicate the activation of ineffective erythropoiesis, which is a predictor of the development of anemia. In this regard, selective prophylaxis of iron deficiency by prescribing a vitamin-mineral complex (Vitrum Prenatal) in the daytime and 100 mg of elemental iron (Sorbifer Durules) in the evening was started [Serov V.P., Burlev V.A. , Konovodova E.N. Treatment of manifest iron deficiency in pregnant women and puerperas (medical technology). M .: MedExpertPress, 2010. - 28 s]. Laboratory study after 3 weeks: red blood cells 3.6 × 10 ¹² / l, HB - 128 g / l, serum iron 20.3 μmol / l, a population of low-activity red blood cells - 17.2%. The obtained indicators indicate a decrease in the proportion of ineffective erythropoiesis, which determines the positive effect of the therapy. A patient has no anemia.

Childbirth ended on time, without complications. A live full-term baby was born weighing 3800 g, 54 cm tall, with a Hangar rating of 8/8 points.

The proposed method for predicting and early diagnosis of anemia is implemented on the basis of the GBUZ TO “Perinatal Center” of the city of Tyumen. Under observation were 79 patients. Table 1 presents the results of the surveys.

The proposed method is easily reproducible, does not require large material costs, allows you to predict the development of the disease, reduces the time of laboratory studies in comparison with the prototype and can be performed in medical institutions. The proposed method allows to predict anemia in pregnant women in 90% of cases. The overall economic effect on the examination of 100 patients compared to the prototype is 11370 rubles.

A method for predicting anemia in pregnant women

Table 1 The results of the prognosis of anemia in pregnant women by the proposed method Group number of pregnant women examined The number of pregnant women examined The population of low-activity red blood cells in peripheral blood
(at %) The development of anemia in pregnant women
(at %) one 10 10-15 2 2 fifteen 16-24 5 3 25 25-30 80 four 25 31-35 90 5 25 36-40 one hundred

Claims (1)

A method for predicting anemia in pregnant women, characterized in that when the number of red blood cells containing no more than 5 granules of formazan in one red blood cell increases in peripheral blood by 25-40%, the development of anemia is predicted by the cytochemical determination of glucose-6-phosphate dehydrogenase activity.