Aug 2026· Archives of Medical Research· Vol 57 8, pp.
103505
· 0 citations· 85 references
Medicine
Abstract
In this narrative review, we outline the historical progression of, and modern advancements in, prenatal diagnosis for monogenic diseases. Initially dependent on invasive sampling procedures, prenatal diagnosis has evolved through the integration of high-throughput molecular techniques, such as next-generation sequencing. These techniques now allow for the comprehensive detection of pathogenic variants across the genome. A major breakthrough occurred with the discovery of cell-free fetal DNA (cfDNA) in maternal plasma, originating from the placenta. This discovery enabled the development of non-invasive prenatal testing (NIPT), which is now widely used. However, non-invasive prenatal diagnosis (NIPD) for monogenic diseases took longer to develop, mainly due to the need for highly sensitive sequencing methods and sophisticated statistical analysis to determine maternal transmission. Currently, only a few countries have implemented NIPD in clinical settings. The multitude of available technologies and protocols can complicate the information provided during antenatal consultations. However, mastery of the knowledge and ethical issues surrounding NIPD will ensure optimal service and better care for pregnancies at risk of monogenic disease transmission.
A structured narrative review of advances in molecular diagnostic technologies across the preconception, preimplantation, and prenatal stages over the past five years is provided, aiming to enhance resolution while balancing health-economic considerations and ethical standards.
De-Yuan Kong, Jia-Ning Zhao, Haichang Diao et al.· Current Issues in Molecular...· 0 citations
OBJECTIVE
Spontaneous abortion is the most common complication in early pregnancy, with chromosomal abnormalities accounting for up to 50% of first-trimester losses. Conventional genetic analyses often rely on invasive procedures that may be limited by technical challenges and patient reluctance. This study aimed to evaluate the feasibility and diagnostic performance of cell-free DNA (cfDNA) based testing as a non-invasive approach for detecting chromosomal abnormalities in first-trimester spontaneous abortion.
METHODS
This study included pregnant individuals diagnosed with spontaneous abortion between 7+0 and 14+6 weeks of gestation from 2022 to 2025. cfDNA was extracted from maternal plasma and analyzed using the VeriSeq™ NIPT Solution v2 (Illumina). A subset of participants who underwent dilation and curettage (D&C) also had fetal tissue analyzed via chromosomal microarray (CMA), which served as the reference standard. Primary outcomes included the rate of non-informative results (NIR), fetal fraction (FF) ≥4%, and detection rate of chromosomal abnormalities. Sensitivity, specificity, and predictive values of cfDNA were calculated relative to CMA.
RESULTS
A total of 299 participants underwent cfDNA testing, of whom 28 also had fetal tissue CMA. The mean FF was 5.83%, with 231/299 (77.3%) samples having FF ≥4%. NIR occurred in 6 cases (2.0%), while 62 (20.7%) had FF <4%. Among cfDNA results with FF ≥4%, 130/231 (56.3%) showed chromosomal abnormalities, with trisomy 16 being the most common. In the D&C subgroup, cfDNA and CMA results were fully concordant (sensitivity and specificity 100%). FF was positively correlated with gestational age (r = 0.42, p < 0.001).
CONCLUSIONS
cfDNA-based testing represents a feasible non-invasive method for detecting chromosomal abnormalities following first-trimester pregnancy loss. It may offer an effective alternative to invasive genetic testing, particularly when tissue sampling is unavailable.
Giovanni Savarese, Maurizio Guida, Monica Ianniello et al.· Fetal Diagnosis and Therapy· 0 citations
We read with great interest the European Hematology Association (EHA) recommendations by de Montalembert et al. on preconceptual and antenatal screening and prenatal diagnosis for hemoglobino-pathies. The manuscript provides a timely and valuable framework for genetic counseling and reproductive decision ‐ making in thalassemia and sickle cell disease, particularly by emphasizing non ‐ directive counseling and the need to adapt diagnostic strategies to local healthcare infrastructures. 1 We would like to comment specifically on Question 4, concerning the laboratory tests that should be used for prenatal diagnosis. The authors appropriately state that invasive prenatal diagnosis based on chorionic villus sampling or amniocentesis remains the most established diagnostic approach, and they also highlight the current limitations of non ‐ invasive prenatal testing (NIPT)/diagnosis for hemoglobinopathies. In particular, they cite previous data showing that the performance of non ‐ invasive approaches for sickle cell disease has not yet reached that of invasive prenatal diagnosis. 1 However, we believe that this section may benefit from a more explicit distinction in light of recent advances in non ‐ invasive prenatal diagnosis (NIPD) based on cell ‐ free fetal DNA and digital polymerase chain reaction (PCR). Our group has recently published a pilot study evaluating third ‐ generation digital PCR for NIPD of sickle cell disease using relative mutation dosage analysis in maternal plasma. In this study, nine pregnant women carrying the HbS heterozygous variant were included. Z ‐ score classification correctly identified the fetal genotype in all conclusive cases, with one inconclusive result, and postnatal confirmation showed 100% concordance
P. Ropero, F. González, Miguel Gómez et al.· HemaSphere· 0 citations
BACKGROUND
C-MoKa (Chromosome Conformation-based Karyotyping) is a novel 3D genome mapping platform, that enables simultaneous detection of structural variations (SVs), aneuploidies, copy number variations (CNVs), and uniparental disomy (UPD) in a single test. However, its performance in routine prenatal diagnosis is still unexplored. This study aimed to comprehensively evaluate the diagnostic performance and clinical utility of C-MoKa in prenatal diagnosis, and to assess its technical concordance with standard of care (SOC) testings in prenatal diagnosis.
METHODS
A two-phase study was designed. In phase 1, 56 retrospective participants with known chromosomal abnormalities (CAs) were recruited, and C-MoKa was performed on their cultured amniotic fluid (AF) samples. Concordance between C-MoKa and known CAs was analyzed. In phase 2, a prospective cohort of 208 participants for prenatal diagnosis were recruited. Samples underwent C-MoKa and karyotyping (KT), with parallel chromosomal microarray analysis (CMA) or improved whole-exome sequencing (iWES). In our study, supplementary copy number probes were enhanced in iWES detection, which could provide ~ 100 kb resolution across the genome, thus either CMA or iWES platform employed was used to assess the CNV detection. Diagnostic yields and concordance were evaluated, and the discordance of SVs and CNVs were further validated by fluorescence in situ hybridization (FISH) and CNV-seq, respectively.
RESULTS
In the retrospective cohort with known CAs, C-MoKa achieved a 94.6% (53/56) diagnostic yield and 92.8% (52/56) concordance with KT + CNV. In the prospective cohort, its diagnostic yield was 21.2% (44/208), higher than KT (12.5%, 26/208) and CNV (18.3%, 38/208). The diagnostic yield (22.6%, 47/208) was achieved when CNV platform and C-MoKa were used together. C-MoKa exhibited concordance rates of 89.4% (186/208) with KT, 89.9% (187/208) with CNV, and 90.4% (188/208) with KT + CNV. Two SVs identified by C-MoKa but missed by KT were successfully determined by FISH, and five samples with additional CNVs identified by C-MoKa were consistently detected by CNV-seq.
CONCLUSIONS
Our findings demonstrate that C-MoKa is a highly effective and reliable method for prenatal diagnosis, exhibiting high concordance with SOC techniques. Compared to the conventional application of "KT + CNV" in prenatal setting, the combined use of CNV and C-MoKa appears to maximize the diagnostic yield.
Ying Zhou, Min Xie, L. Tian et al.· Journal of Translational Med...· 0 citations
Objective To investigate the causes of false-negative results in noninvasive prenatal testing (NIPT) and provide insights for technical optimization and genetic counseling in clinical practice. Methods We retrospectively analyzed clinical and genetic data from three cases with false-negative NIPT results. In case 1, amniocentesis was performed due to fetal growth restriction (FGR) and a single umbilical artery (SUA). In case 2, a female infant was delivered at 35 weeks’ gestation for FGR and vasa previa, with postnatal manifestations including neonatal respiratory distress syndrome, low birth weight, and hexadactyly. In case 3, amniocentesis was carried out for a high-risk NIPT result indicating a 14q duplication. Genetic analyses including karyotyping, copy number variation sequencing (CNV-seq), trio whole-exome sequencing (WES-trio), and fluorescence in situ hybridization (FISH) were performed on amniotic fluid, placental tissue, maternal peripheral blood, and buccal mucosal samples, respectively. Results Ring chromosome 21, dicentric chromosome 18, and multiple chromosomal rearrangements were identified in the three cases. In Case 1, amniotic fluid analysis revealed mosaic r (21) by karyotyping and a terminal deletion of 21q by CNV-seq, which was attributed to dynamic mosaicism and potential cell culture artifacts. In Case 2, karyotyping and WES detected 42% and 65% mosaic dic (18) in peripheral blood, respectively, whereas FISH on buccal mucosal cells only identified 8% mosaicism. This substantial discrepancy reflected tissue-specific mosaic distribution between buccal cells (ectoderm) and peripheral blood (mesoderm) of dic (18). In Case 3, prenatal karyotyping and CNV-seq confirmed a pathogenic 5p deletion in the fetus. Subsequent genetic testing of placental specimens revealed complex mosaic CNVs (10%–52%), whereas umbilical cord findings were consistent with amniotic fluid results. Complex chromosomal rearrangements occurring during early embryogenesis led to widespread multi-CNV mosaicism across placental and fetal tissues. Such heterogeneous genomic alterations are incompletely represented in cffDNA, resulting in concurrent false-positive and false-negative NIPT results. Conclusion Mosaicism levels, tissue distribution variability, cellular heterogeneity, and complex chromosomal structural abnormalities are key contributors to false-negative NIPT results. Our findings highlight the importance of comprehensive clinical and genetic evaluation in managing discordant NIPT findings.
Ning Huang, Yonghua Xu, S. Ding et al.· Frontiers in Genetics· 0 citations
Circulating fetoplacental nucleic acids have transformed prenatal medicine within a single generation, and screening based on cell-free DNA (cfDNA) is now offered routinely in many health systems. The field is moving quickly from the detection of whole-chromosome aneuploidy towards earlier sampling, sub-chromosomal resolution, monogenic diagnosis and the interrogation of epigenetic marks, yet the evidence supporting these extensions is uneven and the downstream consequences for children are rarely examined. This critical narrative review evaluates the state of knowledge on early prenatal detection of fetal genetic variants and epigenetic alterations through maternal blood sampling, and appraises the paediatric implications of an expanding prenatal detection frontier. Literature was identified through structured searching of Europe PMC and MEDLINE, Crossref Metadata Search, OpenAlex, Semantic Scholar and targeted retrieval of professional society statements, supplemented by backward and forward citation tracking. Evidence was appraised for design adequacy, confirmatory testing, spectrum of enrolled participants, and separation of analytical from clinical validity. Three findings dominate the synthesis. First, diagnostic confidence declines sharply and predictably as the target moves from common autosomal trisomies to rare autosomal trisomies, copy number variants and single-gene conditions, and this gradient is driven more by target prevalence and by placental biology than by sequencing chemistry. Second, DNA methylation currently functions far more securely as an analytical instrument, supporting fractional quantification and tissue-of-origin deconvolution, than as a validated diagnostic target for fetal disease, and the developmental literature that motivates epigenetic prediction rests overwhelmingly on postnatal tissues rather than on prenatal plasma. Third, paediatric evidence is the weakest link in the chain: prenatal detection demonstrably alters the ascertainment and the age distribution of childhood diagnoses, but longitudinal outcome data for prenatally ascertained children remain scarce. Priorities include phenotype-linked birth cohorts of prenatally screened pregnancies, prospective validation of methylation-based classifiers against paediatric endpoints, and evaluation frameworks that treat placental discordance as clinical information rather than analytical noise.
S. Bittmann, E. Luchter, E. Moschüring-Alieva· Asian Journal of Pediatric R...· 0 citations