Targeted gene panel sequencing for molecular diagnosis of congenital adrenal hyperplasia
Targeted gene panel sequencing for molecular diagnosis of congenital adrenal hyperplasia
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靶向基因组测序用于先天性肾上腺增生的分子诊断
DOI：
10.1016/j.jsbmb.2021.105899
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发表时间：
2021-05-06
影响因子：
4.1
通讯作者：
Ning, Guang
中科院分区：
文献类型：
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作者：
Wang, Wencui;Han, Rulai;Ning, Guang
背景：先天性肾上腺皮质增生症（CAH）是一组常染色体隐性遗传性疾病，由编码类固醇生成酶和辅因子的9个基因发生遗传缺陷所致。
目的：建立针对全部9个CAH候选基因的靶向二代测序（NGS）检测方法。
方法：我们开发了一种针对CAH候选基因（CYP21A2、CYP17A1、CYP11B1、StAR、CYP11A1、POR、HSD3B2、H6PD、CYP11B2）的定制靶向NGS检测方法，并将该检测方法与CYP21A2的多重连接依赖探针扩增技术（MLPA）相结合，应用于469例有CAH样体征和症状的患者。
结果：我们在8个基因中共鉴定出125个变异，涵盖7种变异类型。变异类型包括错义变异（46.8%）、剪接变异（21.5%）、小的插入缺失变异（12.5%）、大的结构变异（11.8%）、无义变异（4.1%）、非翻译区（UTR）变异（2.9%）、同义变异（0.3%）。98.5%（336/341）的病例成功进行了基因分型，即鉴定出双等位基因致病性或可能致病性变异，其中CYP21A2双等位基因变异（n = 254）、CYP17A1（n = 45）、CYP11B1（n = 23）、StAR（n = 7）、HSD3B2（n = 4）、POR（n = 1）、CYP11A1（n = 1）和CYP11B2（n = 1）基因。重要的是，该检测方法发现1例CYP11B1缺陷患者、1例非经典型POR缺陷患者以及2例非经典型CYP17A1缺陷患者，而这些患者的临床诊断结果与之不同。
结论：我们基于NGS的检测方法联合CYP21A2的MLPA技术是对CAH所有亚型进行基因分型的有效工具。该检测在98.5%临床确诊为CAH的患者中成功实现了基因分型。它还有效地助力了罕见亚型以及非经典表型CAH患者的诊断。
Context: Congenital adrenal hyperplasia (CAH) is a group of autosomal recessive genetic diseases caused by genetic deficiency in nine genes encoding steroidogenesis enzymes and cofactors.Objective: To establish a targeted next-generation sequencing (NGS) assay for all nine CAH candidate genes. Methods: We developed a customized targeted NGS assay of CAH candidate genes (CYP21A2, CYP17A1, CYP11B1, StAR, CYP11A1, POR, HSD3B2, H6PD, CYP11B2) and apply this assay plus MLPA of CYP21A2 in a total of 469 patients with CAH like signs and symptoms.Results: We totally identified 125 variants with seven variant types in eight genes. Variant types included missense variant (46.8 %), splicing variant (21.5 %), small indel (12.5 %), large structure variation (11.8 %), nonsense variant (4.1 %), UTR variant (2.9 %), synonymous variant (0.3 %). Successful genotyping, defined as biallelic pathogenic or likely pathogenic variants, was achieved in 98.5 % (336/341) of cases, including biallelic variants in CYP21A2 (n = 254), CYP17A1 (n = 45), CYP11B1 (n = 23), StAR (n = 7), HSD3B2 (n = 4), POR (n = 1), CYP11A1 (n = 1) and CYP11B2 (n = 1) gene. Importantly, the assay found one patient with CYP11B1 deficiency, one patient with non-classic POR deficiency and two patients with non-classic CYP17A1 deficiency while clinically diagnosed differently.Conclusions: Our NGS-based assay plus MLPA of CYP21A2 is a useful tool to genotype all subtypes of CAH. The test successfully achieved genotype in 98.5 % of patients with clinically determined CAH. It also efficiently facilitated the diagnosis of CAH in patients with rare subtypes as well as non-classic phenotypes.