RU2103686C1 - Method to predict the result of pregnancy in women with precarious miscarriage at early terms - Google Patents

Abstract

FIELD: medicine, obstetrics. SUBSTANCE: method deals with quantitative determining placental alpha-2-microglobulin in husband's sperm and at its content value equal or lower 8 mcg/ml spontaneous miscarriage is predicted and at its spermatic level of 16-64 mcg/ml further flow of pregnancy is diagnosed. EFFECT: higher accuracy of prediction being 95%, higher efficiency in controlling women with precarious miscarriage at early terms of pregnancy.

Description

 The invention relates to medicine, namely obstetrics and gynecology, can be used to predict the outcome of pregnancy in women with a threatening miscarriage of early terms (up to 12 weeks) in the sperm of the father of the child.

 The problem of miscarriage remains one of the most urgent in modern obstetrics, including threatening miscarriages of early terms from 13 to 30% according to domestic and from 60 to 78% according to foreign authors.

 As a prototype, a method for predicting the outcome of pregnancy in women with a threatening early miscarriage was selected by examining a specific immunochemical parameter in the blood serum of pregnant women, chorionic alpha globulin (CAH), followed by predicting a miscarriage in the absence of CAH in the blood serum according to a qualitative reaction with an accuracy of up to 88, 9% and with a positive reaction to CAH in the test sample, pregnancy is predicted to persist until the date of delivery with an accuracy of 82.6%.

The disadvantages of the prototype method are as follows:
1. The test for the study of CAH is currently not performed by any laboratory in the country and is not produced either by domestic or foreign firms, therefore its use is limited to copyright sets.

 2. Invasiveness of the method - for the execution it is necessary to study the venous blood of a pregnant woman, and this is a high risk of a woman becoming infected with the AIDS virus.

 These disadvantages can be eliminated in the proposed method.

 The aim of the invention is to predict the outcome of pregnancy in women with a threatening miscarriage in the early stages (up to 12 weeks) for husband's sperm.

 The goal is achieved by examining the content of PAMG-2 (placental alpha-2 microglobulin) in the husband's sperm, and with its level less than or equal to 8 μg / ml, an unfavorable pregnancy outcome is predicted in 95% of cases.

 The novelty of the invention lies in the fact that for the first time it is proposed to predict the outcome of pregnancy in women with the threat of interruption of the early stages by determining the PAMG-2 protein in the sperm of the husband. Earlier studies of husband's sperm on PAMG-2 in women with a threatened miscarriage in order to predict the outcome of pregnancy were not conducted.

 Placental alpha-2 microglobulin (PAMG-2) was first found in aqueous extracts of human placenta in 1976 by D. D. Petrunin et al. And was named organ-specific alpha-2 placental globulin. The new protein was identified in the placenta at all stages of the gestational process, in amniotic fluid, in the blood serum of women, extracts of chorionepithelioma. Subsequently, given the low molecular weight of the protein, it was called placental alpha-2 microglobulin. Later, after the discovery of PAMG-2 in the organs of the reproductive system of men and women, it began to be designated as alpha-2 microglobulin fertility.

 In previous studies, the association of PAMG-2 with fertility in women and men was identified, on the basis of which new methods were developed for determining male fertility, diagnosing luteal phase insufficiency, and predicting gross congenital malformations in the fetus.

 The method is as follows: the study of PAMG-2 is carried out by double immunodiffusion method in agar with a standard test system against PAMG-2. Anti-PAMG-2 antiserum is obtained by immunization of rabbits with a retro-placental blood fraction. The method is semi-quantitative. Before the analysis, a series of dilutions of the test sperm in distilled water in the wells of the polyethylene matrix is prepared, respectively 1: 1, 1: 2, 1: 4, 1: 8, 1:16, 1:32, 1:64. The reaction is carried out on defatted glass slides, filled with molten 1% solution of Difco type agar with 25% sodium chloride solution at the rate of 5 ml of gel per analysis. When the agar hardens, the holes are punched with a special standard stamp "seven". Two "sevens" are made on one glass. Agar is removed from the wells with a needle. Using a Pasteur pipette, the antiserum against PAMG-2 is poured into the central hole of each of the “sevens”, the solution of standard antigen (abortion blood in standard dilution) is placed in the two opposite ones, and the test sperm in different dilutions is used in the remaining four. Glasses are placed in Petri dishes with a moist environment. Immunological manifestation lasts 18-24 hours. The results are read the next day in transmitted light. The last bend of the precipitation line near the well with the appropriate dilution of the studied sperm is taken into account. Thus, a semi-quantitative assessment of the result is obtained. This dilution is multiplied by the sensitivity of the method - 2 μg / ml. The obtained result is compared with the normative indicators of PAMG-2 sperm of men with undisturbed reproductive function. When stating the reaction for PAMG-2 for one analysis, one glass slide with agar is used.

 Common to the prototype method and the claimed method is the study of biological material for protein immunochemical parameter (HAG in the prototype method and PAMG-2 in the claimed) by immunodiffusion analysis with the corresponding standard test systems.

Significant differences are:
firstly, a new diagnostic parameter for husband’s sperm is proposed - PAMG-2, equal to or less than 8 μg / ml for predicting the outcome of pregnancy in women with a threatening early miscarriage.

 secondly, another biological medium is used - husband’s sperm (the prototype method uses the blood serum of pregnant women for the study of CAH).

The invention is illustrated by the following examples:
Example 1. Freedom S., 25 years old, with a history of one spontaneous miscarriage in the early stages, was admitted to the branch of the maternity hospital in the department of pathology of pregnant women with a diagnosis of Pregnancy 8 weeks, the threat of interruption. Pregnancy is desirable. Upon admission, she complained of pain in the lower abdomen, meager alternating discharge from the genital tract. Inspection data at admission: cervix without defects, external pharynx closed. The uterus is enlarged to an 8-week gestational age, a soft consistency, painless, when examined, the uterus is excitable. Appendages are not determined. Discharges are mucous-sacred, scanty. Colposocytology data: surface cells - 0, intermediate - 25, scaphoid - 75 (these indicators made it possible to predict a favorable pregnancy outcome). Examination by the prototype method: a positive reaction to CAH was revealed, which made it possible to predict the continuation of pregnancy. When examining the claimed method: PAMG-2 in the husband's sperm was found at a concentration of 1: 1, which is 2 μg / ml, these data allowed to predict a miscarriage. The woman underwent maintenance therapy (antispasmodic drugs, sulfate magnesia, vitamin therapy, physiotherapy). Two weeks after the start of treatment, a miscarriage occurred. Histological examination of scraping from the uterine cavity indicated the presence of a fetal egg. Thus, the prognosis of miscarriage by the claimed method was confirmed.

 Example 2. Patient 3., 24 years old, re-pregnant, having a history of three spontaneous miscarriages in the early stages, was admitted to the maternity hospital with a diagnosis of Pregnancy 9 weeks, threatening miscarriage. On admission, she complained of sacral discharge from the genital tract for two days, aching pain in the lower abdomen. On examination: the cervix is defective, the external pharynx is closed. The uterus is enlarged to a 9-week gestational age, the tone is increased, and when examined, the uterus is excitable. Appendages are not determined. The excreta are vernal, insignificant. Colposocytology data: surface cells - 0, intermediate - 20, scaphoid - 80, which allows to predict a favorable pregnancy outcome. Examination by the prototype method: CAH in the blood serum is determined (positive reaction). Preservation of pregnancy was predicted. The patient underwent symptomatic therapy aimed at maintaining pregnancy. When examining the claimed method: PAMG-2 in the husband's sperm was found at a concentration of 1: 4 which is 8 μg / ml. An adverse pregnancy outcome was predicted. Three weeks later, despite ongoing maintenance therapy, a miscarriage occurred. Histological examination of scraping from the uterine cavity indicated the presence of elements of the ovum. Thus, the prognosis of miscarriage by the claimed method was confirmed.

 Example 3. Patient N., 23 years old, with a history of one spontaneous miscarriage in the early stages, was admitted to the department of pathology of pregnant women in the maternity hospital with a diagnosis of Pregnancy II weeks, the threat of interruption. Pregnancy from a second marriage, desirable. Upon admission, she complained of pulling pains in the lower abdomen, smearing sacral discharge from the genital tract during the day. On examination: the cervix is conical in shape, free of defects, clean. The uterus is enlarged until the 2nd week of pregnancy is softened, when examined it is excitable. Appendages are not determined. Discharges are vernacular, scarce. Inspection data for colpocytology: surface cells - 0, intermediate - 18, scaphoid - 82, which allows us to predict a favorable pregnancy outcome. Examination data by the prototype method: CAH in the blood serum is determined (positive reaction). Preservation of pregnancy was predicted. When examining the claimed method: PAMG-2 in the husband's sperm was found at a concentration of 1: 8, which is 16 μg / ml, which allows to predict a favorable pregnancy outcome. Freedom was carried out symptomatic therapy aimed at maintaining pregnancy. A month later, patient I. was discharged from the hospital of the maternity hospital with a progressive pregnancy, and a timely delivery occurred at 39 weeks, a girl was born, weighing 3750 g, body length 52 cm, with an Apgar score of 8 - 9 points. After 5 days, the mother and child, in satisfactory condition, were discharged from the maternity hospital home. Thus, the prognosis of labor by the claimed method was confirmed.

 The advantages of the proposed method.

 1. High accuracy of the method - 95%.

 2. Absolute safety and accessibility of the method - associated with the study of husband's sperm at PAMG-2, since there is no need to take blood from a pregnant woman and any other trauma.

 3. Allows you to decide early on the advisability of continuing or terminating a pregnancy from the time of its diagnosis (2 to 3 weeks).

 4. The simplicity and accessibility of using test systems at PAMG-2 make it possible to widely use it in practical health care.

 5. Test systems for PAMG-2 are produced by several diagnostic companies that have the right to implement. At the moment, test systems for PAMG-2 can be purchased at the address: Moscow., LLP., "Rus 2".

 Thus, the claimed method for predicting the outcome of pregnancy in women with a threatening miscarriage of early terms (up to 12 weeks) provides, along with high accuracy and ease of implementation, complete safety for women and the availability of use compared to the prototype method. Using the proposed method will allow you to choose the right tactics for women with a threatening miscarriage in the early stages, which will help reduce the frequency of miscarriage, reduce the economic cost of treatment.

Claims (1)

 A method for predicting the outcome of pregnancy in women with a threatening miscarriage of early terms by quantitative determination of placental alpha-2 microglobulin (PAMG-2), characterized in that PAMG-2 is examined in the sperm of the child’s father and, with its content less than or equal to 8 μg / ml, spontaneous miscarriage.