Background: Next-generation sequencing (NGS), particularly whole-exome sequencing (WES), has become a powerful diagnostic tool for rare genetic conditions. However, its success rate varies based on the underlying genetic etiology and the population studied. Methods: This retrospective study evaluated the diagnostic yield of NGS in a cohort of 137 pediatric patients with suspected rare genetic disorders in Bulgaria, a setting where such testing is not reimbursed and must be self-funded. The patients underwent either WES or targeted gene panel testing based on clinical presentation, family history, and genetic evaluation. Results: The overall diagnostic yield was 45.99%, with WES achieving 51.25% and targeted testing achieving 38.60%. The highest yield was observed in patients presenting with both dysmorphic features and neurodevelopmental delays (62.5%), while the lowest was observed among those with isolated neurodevelopmental issues (10%). A significant portion of the identified variants (35.9%) were novel. Eight patients were diagnosed with copy number variants (CNVs) detected only through WES. Conclusions: Our findings illustrate the value of WES as a first-line test and highlight the impact of deep phenotyping on diagnostic success. This study also emphasizes the need for a population-specific reference genome and equal access to genomic diagnostics in all European countries.

Hereditary tumor predisposition syndromes (HTPSs) significantly increase the risk of developing various cancers, often at earlier ages than seen in the general population. The development and application of next-generation sequencing (NGS) has revolutionized the identification of individuals with HTPS, facilitating early diagnosis, personalized risk assessment, and tailored preventive strategies. However, the widespread implementation of genetic testing for HTPS presents complex ethical and psychosocial issues. This paper examines key ethical considerations surrounding genetic testing for HTPS, including the following: the distinct nature of genetic information and its implications for families; the challenges of informed consent amidst evolving genetic knowledge and direct-to-consumer testing; the complexities of predictive and presymptomatic testing, particularly in minors; and the implications of incidental findings. It further explores the critical issue of genetic discrimination, particularly concerning insurance, employment, and social stigmatization. This paper highlights the importance of balancing individual rights, such as autonomy and privacy, with familial responsibilities and the potential benefits of early detection and intervention. It also underscores the need for robust legal frameworks, comprehensive genetic counseling, and ongoing public education to address the ethical and psychosocial challenges associated with genetic testing for HTPS, with the ultimate goal of maximizing the benefits of genomic medicine while minimizing potential harms.

Acute myelogenous leukemia is a multi-step hematological malignancy, affecting function, growth, proliferation and cell cycle of myeloid precursors. Overall assessment of patients with the disease requires among everything else, a comprehensive investigation of the genetic basis through various methods such as cytogenetic and molecular-genetic ones. This clarification provides diagnostic refinement and carries prognostic and predictive value in respect of essential therapeutic choices. With this review of the literature, we focus on summarizing the latest recommendations and preferred genetic methods, as well as on emphasizing on their general benefits and limitations. Since none of these methods is actually totipotent, we also aim to shed light over the often-difficult choice of appropriate genetic analyses.

The genetic diversity of acute myeloid leukemia creates a major obstacle for current research and clinical practice. Despite advances in molecular genetic techniques, various omics approaches, and artificial intelligence, developing a universal algorithm to thoroughly assess each clinical case remains difficult. Starting with current recommendations and classifications, this narrative review highlights the most important diagnostic options available today, new opportunities that are emerging, and the challenges in diagnosing and managing this complex disease.

Introduction: This study aimed to assess the effectiveness of multiplex ligase-dependent probe amplification (MLPA) in the initial genetic screening of patients with acute myeloid leukemia (AML) since current risk stratification and clinical management depend on molecular-genetic markers. Methods: We performed a prospective case-control study on newly diagnosed patients from the Clinical hematology clinic of UMHAT “St. Marina”, Varna, for the period 02.2022 – 02.2023. MLPA – a semiquantitative PCR-based method, was implemented with probes for 40 AML/myelodysplastic syndrome-typical genetic changes. We compared these findings with a parallelly carried out cytogenetic analysis, part of the routine diagnostic process. Results: We assessed 61 patients – 29 females and 32 males, median age of 61 years for females and 65 for males (min-max 20-89). 34 (56%) of all showed pathological results, while the rest 27 (44%) did not. Of the 34, 22 (65%) had a single gene variant in genes NPM1, DNMT3A, FLT3, and IDH2, isolated or in combination. 18 (53%) of the same 34 also had copy number aberration (CNA) in chromosomes 4, 5, 6, 7, 11, 14, 17, and 21. The latter were either isolated or in combination with other findings. 8 of the 18 cases also underwent cytogenetic analysis, with concordance between the two methods in 4. Conclusion: MLPA is an informative method for initial genetic assessment in addition to cytogenetic analysis. Still, more patients are needed to draw finite conclusions on its eligibility for routine use. Given the significant percentage of normal results – 44%, simultaneous evaluation of more genetic markers, included in current guidelines, is reasonable.

Background and Clinical Significance: Ciliopathies are a heterogeneous group of diseases caused by damage to the primary cilium. Disorders of ciliary motility can lead to a wide range of clinical manifestations, including infertility, lateralization defects, lung infections, and more. Some ciliopathies associated with kidney disease include nephronophthisis, polycystic disease, and renal cell carcinoma. Since they are clinically and genetically diverse, their diagnosis may require a longer time and one or more genetic assays. Case presentation: We present the case of a 43-year-old man with a wide anamnesis, including unexplained nephrolithiasis, bronchiectasis, recurrent otitis media since infancy, appendicular lithiasis, and infertility. After a long history of various clinical examinations and consultations with diverse specialists, he was referred to genetic counseling. Whole exome sequencing (WES) revealed a homozygous pathogenic variant in the RSPH3 gene—NM_031924.8:c.205-2A>G—which was later confirmed through Sanger sequencing. It is classified as pathogenic in widely used databases and is associated with primary ciliary dyskinesia. This condition can present nontypically, and the patients might suffer from an extensive diagnostic odyssey. Being mindful of its clinical and genetic heterogeneity can shorten the period until diagnosis. Conclusions: It is essential to have this condition included in differential diagnosis and involve specialists from the medical/clinical genetic department in a multidisciplinary team. Genetic confirmation through WES or another molecular genetic method is crucial for the therapeutic approach and to adequately perform genetic counseling for patients and their families.