■由于人群的高度遗传异质性,囊性纤维化(CF)的分子诊断在墨西哥具有挑战性。迄今为止,已经报道了46种致病变异(PV),检出率为77%。我们更新了墨西哥患者中导致这种疾病的PV的频谱和频率。
■我们从297例CF患者及其父母的外周血淋巴细胞中提取了基因组DNA。首先,我们使用PCR介导的定点诱变分析了墨西哥人群中5种最常见的PV.在至少有一个确定的等位基因的患者中,使用下一代测序工具和多重连接依赖性探针扩增进行CFTR测序。对于以前未归类为致病性的变体,我们使用了计算机预测的组合,CFTR建模,和临床特征以确定基因型-表型相关性。
■我们确定了95个PVs,检出率提高到87.04%。最常见的变异是p。(PheF508del)(42.7%),其次是p.(Gly542*)(5.6%),p.(Ser945Leu)(2.9%),p.(Trp1204*)和p.(Ser549Asn)(2.5%),和CFTRdel25-26和p.(Asn386Ilefs*3)(2.3%)。其余变异的频率<2.0%,有些是一个家庭的专属。我们确定了10个位于不同外显子的新PV(频率范围:0.1-0.8%),所有这些都产生了结构变化,删除,或蛋白质不同结构域的重复,导致功能失调的离子流。使用不同的计算机软件和美国医学遗传学和基因组学学院(ACMG)和分子病理学协会(AMP)标准使我们能够假设所有这些PV都具有致病性,导致严重的表型。
■在高度异质的群体中,需要不同工具的组合来识别负责CF的变体,并能够建立适当的CF诊断策略,预防,和治疗。
UNASSIGNED: Molecular diagnosis of cystic fibrosis (CF) is challenging in Mexico due to the population\'s high genetic heterogeneity. To date, 46 pathogenic variants (PVs) have been reported, yielding a detection rate of 77%. We updated the spectrum and frequency of PVs responsible for this disease in mexican patients.
UNASSIGNED: We extracted genomic DNA from peripheral blood lymphocytes obtained from 297 CF patients and their parents. First, we analyzed the five most frequent PVs in the Mexican population using PCR-mediated site-directed mutagenesis. In patients with at least one identified allele, CFTR sequencing was performed using next-generation sequencing tools and multiplex ligation-dependent probe amplification. For variants not previously classified as pathogenic, we used a combination of in silico prediction, CFTR modeling, and clinical characteristics to determine a genotype-phenotype correlation.
UNASSIGNED: We identified 95 PVs, increasing the detection rate to 87.04%. The most frequent variants were p.(PheF508del) (42.7%), followed by p.(Gly542*) (5.6%), p.(Ser945Leu) (2.9%), p.(Trp1204*) and p.(Ser549Asn) (2.5%), and CFTRdel25-26 and p.(Asn386Ilefs*3) (2.3%). The remaining variants had frequencies of <2.0%, and some were exclusive to one family. We identified 10 novel PVs localized in different exons (frequency range: 0.1-0.8%), all of which produced structural changes, deletions, or duplications in different domains of the protein, resulting in dysfunctional ion flow. The use of different in silico software and American College of Medical Genetics and Genomics (ACMG) and the Association for Molecular Pathology (AMP) criteria allowed us to assume that all of these PVs were pathogenic, causing a severe phenotype.
UNASSIGNED: In a highly heterogeneous population, combinations of different tools are needed to identify the variants responsible for CF and enable the establishment of appropriate strategies for CF diagnosis, prevention, and treatment.