RU2394495C1 - Method for prediction of risk of early fetal loss in ivf induced pregnancies - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to new reproductive technologies in obstetrics. It involves immune-enzyme analysis of a biological fluid. In women with induced pregnancy, the follicular fluid is analysed for specific immunocomplexes, namely alpha 2-macroglobulin-immunoglobulin G (MG-IgG) and lactoferrin immunoglobulin G (LF-IgG), and if MG-IgG is less than 0.6 mcg/ml, and LF-IgG is less than 0.8 mcg/ml, high probability of early miscarriage is predicted.

EFFECT: method allows developing a method for prediction of early miscarriage that allows for well-timed administration of high-level luteal phase support immediately after oocyte sampling and without waiting for IVF program termination.

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Description

The invention relates to medicine, in particular to new reproductive technologies in obstetrics.

The most important component of a person’s general health and quality of life is reproductive health, and one of its main components is the ability to conceive, bear and give birth to children. The loss of this ability is the tragedy of the family and society, and from this perspective, infertility in marriage is a modern medical and social problem that requires further study, especially since in Russia every 4-5th married couple is infertile. The growing interest in the development of assisted reproductive technologies (ART) worldwide is due to the priority issue of infertility. The introduction and modernization of methods of new reproductive technologies contributed to an in-depth study of the reproductive process and significantly expanded the ability to restore reproductive function in the most severe forms of female and male infertility. At the same time, the effectiveness of the implemented in vitro fertilization (IVF) programs does not quite satisfy reproductive medicine specialists: the pregnancy rate after IVF is still low - about 28% for women under 35 years old. The preterm birth rate is 33.6%, and in every 5 women the pregnancy is terminated before 20 weeks of gestation.

Recently, more and more attention has been paid to studying the role of cytokines and specific immunocomplexes in various bio-fluids, which take part in the mechanisms of gestation and the development of pregnancy pathology. Thus, the concentration of certain cytokines in the composition of follicular fluid, according to some authors, reflects the degree of maturity and quality of oocytes. Determination of specific immunocomplexes in the blood of pregnant women allows timely diagnosis of an extremely serious pregnancy complication - eclampsia (Concentration of macroglobulin-IgG complexes as a possible criterion for predicting gestosis and assessing its severity. / V.N. Zorina, PMZorina, V.G. Levchenko and others // Clinical and laboratory diagnostics. - 2001. - No. 8. - S.48-49).

A known method of enzyme immunoassay in the blood serum of the cytokine - IL-1β in the early stages of gestation (11-15 days) during physiological pregnancy. Increased rates of this cytokine correlate with miscarriage in early pregnancy (Fortunato SJ IL-1β is a better inducer of apoptosis in human fetal membranes than IL-6. / SJ Fortunato, R. Menon // Placenta. - 2003. - Vol.24 (10). - P.922-928).

The disadvantages of the method:

1) high individual variability of indicators for levels of a given cytokine in blood serum;

2) studies were conducted during pregnancy, which occurred in natural cycles without stimulation of ovulation, in the first trimester (6-12 weeks) of pregnancy.

There is a method of enzyme immunoassay in the blood serum of the concentration of another cytokine - IFN-γ in the blood of women who participated in IVF programs on the 15th day after the transfer of embryos into the uterine cavity (Maternal serum levels of interferon-gamma and interleukin-2 soluble receptor-alpha predict the outcome of early IVF pregnancies. / SJFasouliotis, SDSpandorfer, SSWitkin et al. // Hum Reprod. 2004. - Jun; 19 (6) .- P.1357-63).

A high concentration of IFN-γ in serum is often accompanied by early reproductive loss.

The disadvantages of the method:

1) high individual variability of indicators;

2) the determination of cytokine (IFN-γ) is carried out in the blood serum of women already in the presence of established pregnancy.

The objective of this invention is to create a sufficiently accurate and informative method for predicting early fetal losses in IVF program participants already on the day of oocyte collection by analyzing the composition of follicular fluid by the enzyme immunoassay.

The problem is achieved by predicting the risk of early fetal loss during induced pregnancy in IVF programs by studying the biological fluid by the enzyme immunoassay. In women, in the event of an induced pregnancy in the follicular fluid, the content of specific immune complexes is determined: alpha 2-macroglobulin - immunoglobulin G (MG-IgG) and lactoferrin - immunoglobulin G (LF-IgG) and with an MG-IgG value of less than 0.6 μg / ml and LF-IgG less than 0.8 μg / ml predict a high probability of an early miscarriage.

The novelty of the invention.

In women, in the event of an induced pregnancy, the content of specific immune complexes of MG-IgG and LF-IgG is determined in the follicular fluid.

When the MG-IgG value is less than 0.6 μg / ml, and LF-IgG less than 0.8 μg / ml, a high probability of an early miscarriage is predicted.

The set of essential features of the invention allows you to create a method for predicting miscarriage in early pregnancy, which allows you to assign intensive support for the luteal phase in a timely manner, immediately after taking the oocytes, without waiting for the end of the IVF program.

These indicators were chosen due to the fact that the above specific complexes have immunoregulatory activity, since they include immunoglobulin G, and the second component is regulatory proteins: alpha 2-macroglobulin (MG) or lactoferrin (LF). In all likelihood, these proteins are synthesized in the follicular fluid and take part in the mechanisms of gestation (“The role of macroglobulins in the reproductive function” (literature review) / N.A. Zorin, V.G. Levchenko, PMZorina et al. // Obstetrics and Gynecology. - 2005. - No. 4. -C.7-9; Birkenmeier G. "Targetting the proteinase inhibitor and immune modulatory function of human alpha 2-macroglobulin" / G. Birkenmeier // Mod. Asp. Immunobiol. - 2001. - Vol.2. - P.32-36; Urdl W. “Criteria of the maturity of follicle and oocyte within the scope of in vitro fertilization.” An overview / W. Urdl // Wien Med Wochenschr. - 1991. - Vol .141 (1-2) .- P.2-9; High concentrations of lactoferrin in the follicular fluid correlate with embryo quality during in vitro fertilization cycles / A. Yanaihara, K. Mitsukawa, S. Iwasaki et al. // Fertil Steril. - 2006. - Vol.6. - P.311-320).

However, in the literature there is no information regarding the use of specific immune complexes as prognostic criteria for the outcome of an induced pregnancy.

The claimed method was applied in 61 participants of the IVF program with analysis of the effectiveness of the programs and the outcome of the pregnancy. In 30 patients, pregnancy in IVF programs did not occur, in 45 patients the program ended with the onset of pregnancy, of which in 6 patients the pregnancy terminated at 7–9 weeks of gestation. The results of the study are presented in table 1.

Table 1 The content of immunocomplexes MG-IgG and LF-IgG (μg / ml) in the follicular fluid of IVF participants. Indicators Groups surveyed Ineffective eco-programs Programs with the onset of pregnancy "ECO +" 1 fruit 2 fruits miscarriage n thirty 34 5 6 MG-IgG (M ± m) 0.87 ± 0.103 0.95 ± 0.19 1.18 ± 0.18 0.47 ± 0.05 * LF-IgG (M ± m) 0.81 ± 0.05 0.98 ± 0.08 1.28 ± 0.16 0.65 ± 0.05 * Notes: * - the difference in indicators is statistically significant compared to the level during normal pregnancy, regardless of the number of fetuses.

When using multivariate analysis, the significance of differences for MG-IgG complexes was 0.019 according to the Kruskal-Wallis Test, and 0.027 according to Van der Waerden. For LF-IgG complexes, 0.013 and 0.01, respectively.

The method is as follows. In women participating in IVF programs, during transvaginal puncture, simultaneously with oocyte collection, follicular fluid is taken and the concentration of specific immune complexes MG-IgG and LF-IgG is determined by the enzyme immunoassay. The results are evaluated as follows: at a level of MG-IgG less than 0.6 μg / ml and LF-IgG less than 0.8 μg / ml, a high risk of early miscarriage in the event of an induced pregnancy is predicted.

Example 1. Pregnant M., 30 years old, Protocol No. IVF E / 596. This is a third pregnancy: the first ended in an urgent delivery (3400.0-51.0), the second - a medical abortion complicated by endometritis. The duration of infertility before joining the IVF program is 7 years. Regarding tubal-peritoneal infertility of inflammatory genesis, IVF was taken into the program, according to a long standard protocol using a-GN-RH, superovulation was stimulated with gonadotropins. At the peak of superovulation induction, 13 oocytes were taken, 10 good quality embryos were obtained. At the stage of oocyte collection in follicular fluid, the concentrations of MG-IgG = 0.4 μg / ml and LF-IgG = 0.5 μg / ml were determined. 2 embryos were transferred to the uterine cavity at the stage of 6 cell division. Therapy, aimed at supporting the luteal phase, was prescribed prophylactic. With confirmation of the fact of pregnancy, therapy was strengthened, and pregnancy progressed until the period of 6-7 weeks. However, subsequently, the pregnant woman was hospitalized in the gynecological department with signs of a threatened miscarriage and subsequently a miscarriage occurred in the period of 7-8 weeks.

Example 2. Patient P.A., 26 years old, protocol No. of conducting the IVF program E / 503. Real pregnancy is the second, the first is a medical abortion, in the period of 12 weeks in connection with the transferred rubella. The duration of infertility before joining the IVF program is 6 years, the diagnosis is verified laparoscopically. Regarding tubal-peritoneal infertility of inflammatory genesis, IVF was taken into the program, according to a long standard protocol using a-GN-RH, superovulation was stimulated with gonadotropins. At the peak of superovulation induction, 20 oocytes were taken, 16 good quality embryos were obtained. At the stage of oocyte collection in the follicular fluid, the concentrations of MG-IgG-1.2 μg / ml and LF-IgG were determined to be 1.45 μg / ml. 3 embryos were transferred to the uterine cavity at the stage of 4-cell division. Therapy, aimed at supporting the luteal phase, was prescribed prophylactic. When pregnancy was confirmed, therapy was reinforced. The further course of pregnancy proceeded without complications and ended in the birth of twins in the period of 38 weeks.

Thus, the proposed method for predicting an early miscarriage in an induced pregnancy by examining the immune complexes in the follicular fluid in women participating in IVF programs allows a timely assessment of the risk and prediction with a high degree of probability of early miscarriage during pregnancy in IVF programs. A timely prognosis and risk assessment of early fetal loss will allow taking measures at the stages of early gestation: to prescribe intensive therapy during the luteal phase support period or to use the cryotransfer of the obtained embryos in another cycle when more favorable conditions are created for implantation and pregnancy development.

Claims (1)

A method for predicting the risk of early fetal loss during induced pregnancy in IVF programs by examining the biological fluid by the enzyme immunoassay, characterized in that in women, in the event of induced pregnancy, follicular fluid determines the content of specific immune complexes alpha 2-macroglobulin - immunoglobulin G (MG-IgG) and lactoferrin - immunoglobulin G (LF-IgG) and when the MG-IgG value is less than 0.6 μg / ml, and LF-IgG less than 0.8 μg / ml predict a high probability of an early miscarriage.