Pacheco-Orozco Rafael Adrián, Devia Angela, Manzi Eliana, Franco Alexis Antonio, Pachajoa Harry, Medina Valencia Diego
Fundación Valle del Lili, Cali, Colombia.
Facultad de Ciencias de la Salud, Universidad Icesi, Cali, Colombia.
Andes Pediatr. 2024 Oct;95(5):525-532. doi: 10.32641/andespediatr.v95i5.5066.
Inherited Bone Marrow Failure syndromes account for approximately 25% of cases of aplastic anemia in pediatric patients. Next-generation sequencing (NGS) technologies have allowed the diagnosis of an increasing number of hereditary causes of bone marrow failure.
To determine the diagnostic yield and clinical concordance of NGS in the diagnosis of a cohort of pediatric patients with bone marrow failure.
Patients included were those aged between 0-17 years with a diagnosis of Bone Marrow Failure Syndrome according to the ICD-10 classification codes, who had undergone a genetic study between 2018 and 2022. The information was obtained from the electronic medical records system. Genomic DNA was isolated and quantified through the Qubit™ 3.0 fluorometer. Regions of interest were selected using a hybridization probe that included the intronic and exonic regions adjacent to the genes included in the panel. Clonal amplification and paired-end sequencing of the selected regions were performed using the Illumina MiSeq™ system. Bioinformatics analysis was performed in alignment with the reference genome (GRCh38). Variants classified as probably pathogenic or pathogenic were confirmed through Sanger sequencing.
Out of 18 patients included, a genetic diagnosis was achieved through NGS in 5 (27.8%) of them: two cases of Fanconi Anemia, two cases of Dyskeratosis Congenita, and one case of TP53- associated bone marrow failure. Clinical concordance was 100%. Two novel variants were found in the FANCA and PARN genes as causing disease.
The use of NGS in patients with bone marrow failure identified the etiology in close to a third of patients of our cohort, with higher yield in patients with a clear clinical diagnosis and syndromic features.
遗传性骨髓衰竭综合征约占小儿再生障碍性贫血病例的25%。新一代测序（NGS）技术使越来越多的遗传性骨髓衰竭病因得以诊断。
确定NGS在一组小儿骨髓衰竭患者诊断中的诊断率及临床一致性。
纳入的患者年龄在0至17岁之间，根据ICD - 10分类编码诊断为骨髓衰竭综合征，于2018年至2022年期间接受了基因研究。信息从电子病历系统获取。通过Qubit™ 3.0荧光计分离并定量基因组DNA。使用包含与该面板中基因相邻的内含子和外显子区域的杂交探针选择感兴趣区域。使用Illumina MiSeq™系统对所选区域进行克隆扩增和双端测序。与参考基因组（GRCh38）比对进行生物信息学分析。通过桑格测序确认分类为可能致病或致病的变异。
在纳入的18例患者中，5例（27.8%）通过NGS获得了基因诊断：2例范可尼贫血、2例先天性角化不良和1例与TP53相关的骨髓衰竭。临床一致性为100%。在FANCA和PARN基因中发现两个导致疾病的新变异。
在骨髓衰竭患者中使用NGS确定了近三分之一队列患者的病因，在临床诊断明确且有综合征特征的患者中诊断率更高。
