The rapid development of genomic technologies, especially chromosomal microarray analysis and new-generation sequencing, has led to significant opportunities for the molecular diagnosis of various mutations/genetic variants. In recent years, there is initiated the widespread using of new methods of genome analysis in various topics of medical genetics, including the evaluation of patients with disorders of sex differentiation, abnormal development of the reproductive system, failures of reproductive function, studies of abortion material of spontaneous interrupted pregnancies, as well as in preimplantation genetic diagnosis. The presented review covers the results of genome research in reproductive genetics and their place in genetic evaluation of patients with reproductive disorders.

We report two familial cases of insertional translocations with chromosomе 2. Both abnormalities were derived from fathers having a balanced insertion. In case 1 patient was referred for evaluation because of developmental delay, seizures of unknown etiology and slight dysmorphic features showed a normal karyotype. Сomplex approach to diagnosis of the proband’s and his family allowed to determine karyotype as 46,XY,der(2)ins(2;7)(q35;q31.33q34)pat. In case 2 patient with neonatal seizures was referred for evaluation. Combined with the conventional cytogenetic studies and FISH analyses proband’s karyotype was determined as 46,XY,rес(2)dup(2q)inv ins(2)(р11.2q23.3q31)pat. In both this cases only complex approach such as CMA, FISH and conventional cytogenetics allows to perform a complete quality diagnosis.

In the presented «case-control» study 890 residents of the Kemerovo Region subject to age, sex, ethnicity and smoking status were included. We formed two groups: 1) «Case» - 440 newly diagnosed lung cancer patients undergoing a medical treatment in the Kemerovo Regional Oncology Center (diagnosis «lung cancer» was determined by experienced doctors from the Kemerovo Regional Oncology based on the results of special medical examination); 2) «Control» - 450 healthy donors of the Kemerovo Regional Center of Blood Transfusion. The aim of this study was the analysis chromosomal aberrations (CAs) in lung cancer patients and healthy donors resident in the same territory. The following methods were used in our investigation: the routine method of lymphocytes cultivation and chromosomal aberration analysis. Statistical analysis were performed using nonparametric statistics (Mann-Whitney U Test for paired comparison of quantitative characteristics), logistic regression. It was determined that the CAs frequency (both chromatid- and chromosome-type aberrations) was significant increased in lung cancer patients compared to control group. Furthermore in 8 lung cancer patients were found Rogue cells that were contained polycentric, ring chromosomes and multiple pair dot fragments.

Cytogenetic methods of cell study of bone marrow and peripheral blood in individuals with onco-hematological diseases are widely used for stating a diagnosis and severity assessment. Diversity of methods of chromosome painting enables to fully assess genetic aberrations that is important for treatment strategy identification and prediction of course and prognosis of the disease. Since laboratories often have funding limits it is important to develop approaches for selecting a cytogenetic method for the analysis. The goal of the present work is to analyze cytogenetic abnormality in bone marrow cells in individuals with chronic leukemia by means of three methods of painting that enabled to form a methodology of cytogenetic research for different groups of diseases (chronic myeloleukemia, chronic lymphocytic leukemia, multiple myeloma and primary myelofibrosis). In the present work we have used: G-banding, 24-colour FISH technique. For the individuals with CLL we have used of locus-specific probes: ATM(11q22)/TP53(17p13) and DLEU(13q14)/LAMP(13q34)/cen12(12p11.1;12q11.1). Summarizing the research results we have come to the following conclusions: differential painting enables to identify all chromosome transformations but only in case of ideal chromosome quality which is not the case with bone marrow. The use of 24-colour FISH technique enables to identify translocations even if the chromosome quality is not good. This method attained credibility for searching complex translocations. The cocktail of locus-specific probes for CLL cases demonstrated high efficiency and a high speed of obtaining results. Cytogenetic analysis in the dynamics allows evaluating the peculiarities of a mutation process in the course of treatment.

Tuberculosis (TB) is a global health problem, both in the world and in our country, especially in the Siberia and the Far East. Well known that there are genetic factors that influence to TB predisposition. The aim of this work was studying the role of IL-12 / IFN-g gene polymorphisms in the development of different clinical forms of TB. The study was conducted in a population of russians in Tomsk: TB patients (n = 331) and healthy donors (n = 279). Patients were divided into subgroups: patients with primary (n = 61) and secondary (n = 270) TB. As well as patients with infiltrative (n = 155) and disseminated (n = 68) TB. The effect of allele A and genotype AA of the STAT1 gene on the development of secondary TB (p = 0.0097) was shown. Also, the association of the C allele of the IL12RB1 gene and the CC genotype with the development of infiltrative TB has been identified. Our results suggest differences in genetic predisposition to various forms of TB.

Phenotypic manifestations of X-chromosome;autosome translocations, in contrast to autosome;autosome translocations, often depend on several factors: the location of the break points on both chromosomes and the features of X-chromosome inactivation. Due to the development of molecular cytogenetic and genetic methods, at the present time we can investigate each specific case of such translocations in details, that allows us better understanding the causes of the pathological phenotype. The aim of the present study was evaluation the effect of the X-chromosome inactivation on the clinical manifestation of various X;autosome translocations. Break points and X-inactivation were assessed by array-CGH (8х60K, Agilent Technologies) and methyl-sensitive PCR at AR gene, espectively. Three cases of X;autosome translocations, the feature of X inactivation, and the clinical picture accompanying chromosomal rearrangement were analyzed. In the case of an unbalanced translocation 46,X,t(X;3)(p11.3;q21.3), the X-chromosome inactivation has a protective effect on the phenotype, whereas second patient with the balanced translocation 46,X,t(X;9)(q22;q13) exhibits severe clinical symptoms, possibly because of partial functional monosomy of the chromosome 9. Furthermore, the phenotype of the patient may be affected by the additional microdeletion that was found by aCGH at the 22q11.22. Finally, in the third case where the distal region of the short arm of the X chromosome is involved in the translocation 46,X,t(X;10)(p22.2;q11.2), the inactivation process is not associated with the patient phenotype, since the Xp22.2 region escape inactivation. For a detailed analysis of the phenotypic manifestations of the X;autosome translocations complex investigation with various molecular diagnostic methods is required, including cytogenetic, molecular cytogenetic methods for analyzing the structure of rearranged chromosomes, and the analysis of the X chromosome inactivation.

Relevance. A comparison of the molecular karyotypes of the cell-free DNA from the blastocoele fluid of the blastocyst, the embryoblast and trophectoderm cells provides new possibilities for studying of the cytogenetic mechanisms of the formation of numerical chromosomal abnormalities at the preimplantation stage of human development. In addition, such analysis allows us to evaluate the diagnostic yield of cell-free DNA as an additional source of information about the embryo karyotype for preimplantation genetic diagnosis. Aim. Comparative molecular cytogenetic karyotyping of embryoblast, trophectoderm and the cell-free DNA from the blastocoele fluid of the blastocyst. Materials and methods. Twenty-nine human blastocysts of the 5th day of development were analyzed by metaphase and array comparative genomic hybridization. Results. Cell-free DNA was successfully amplified in 86.2% (25/29) of the samples. Blastocysts were euploid in 31%, 36% and 28% of cases according to the results of analysis of trophectoderm, embryoblast and сell-free DNA, respectively. Only 3 out of 29 (10.3%) blastocysts had a normal karyotype according to the analysis of all three samples. A total of 175 aneuploidies were detected. Trisomies, monosomies, partial tri- and monosomies were observed at a frequency of 47.4%, 46.9%, 5.1%, and 0.5%, respectively. The prevalence of trisomy was noticed in the embryoblast, which is not available for preimplantation genetic screening. Chromosomal mosaicism was detected in 14 examined blastocysts (48.2%). The reciprocal aneuploidies presented by combination of trisomy and monosomy with one pair of homologous chromosomes were described in 44.8% of blastocysts. A total of 25 reciprocal aneuploidies were observed with involvement of 50 from 175 (28.5%) detected aneuploidies. Conclusions. The cell-free DNA can be successfully amplified and analyzed by current molecular cytogenetic techniques. The results of comparative molecular karyotyping of cell-free DNA, embryoblast and trophectoderm cells indicate an underestimation of the frequency of aneuploid and mosaic blastocysts. It was found that in 72.4% of cases the molecular karyotypes of the embryoblast and trophectoderm are not identical. This finding provides evidence for impossibility of a direct extrapolation of the results of preimplantation genetic screening of trophectoderm cells to the inner cell mass. Accordingly, the cell-free DNA from the blastocoele fluid can be considered as an important additional source of information about the embryo karyotype in preimplantation genetic diagnosis.