Genetic diagnosis of non-classical 21-hydroxylase deficiency by the new nanopore sequencing detection method
10.3760/cma.j.cn114452-20220727-00438
- VernacularTitle:纳米孔测序技术在非经典型21-羟化酶缺乏症基因诊断中的应用
- Author:
Yanjie XIA
1
;
Peng DAI
;
Huikun DUAN
;
Panlai SHI
;
Shanshan GAO
;
Xueyu GUO
;
Ning LIU
;
Xiangdong KONG
Author Information
1. 郑州大学第一附属医院遗传与产前诊断中心，郑州450052
- Keywords:
Nanopore sequencing;
21-Hydroxylase deficiency;
Congenital adrenal hyperplasia;
CYP21A2 gene
- From:
Chinese Journal of Laboratory Medicine
2023;46(1):74-80
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To summarize initial experience of applying nanopore third-generation sequencing detection method (nanopore sequencing) for genetic diagnosis of non-classical 21 hydroxylase deficiency (NC 21-OHD), and to explore its performance and application prospects.Methods:Clinical data of the two NC 21-OHD patients, who were hospitalized at the First Affiliated Hospital of Zhengzhou University in May 2019, were collected. Peripheral venous blood was collected and genome DNA extracted. Genetic variants was detected by nanopore sequencing and underwent bioinformatic analysis. Pathogenetic mutations in CYP21A2 gene were validated with PCR-sanger sequencing in the two patients and their parents.Results:The average reads length and sequence depth in the patient one was 12, 792 bp and 27.19×. The average reads length and sequence depth in the patient two was 13, 123 bp and 21.34×. Compound variants of c.293-13C>G/c.844G>T (p.Val282Leu) and c.332_339delGAGACTAC (p.Gly111Valfs)/c.844G>T (p.Val282Leu) were detected in these two patients, which were consistent with clinical phenotype of NC 21-OHD. Further analysis showed that c.293-13C>G mutation was inherited from her father and c.844G>T (p.Val282Leu) mutation was inherited from her mother for the patient one. The c.844G>T (p.Val282Leu) mutation was inherited from her father and c.332_339delGAGACTAC (p.Gly111Valfs) mutation from her mother.Conclusions:The heterozygous mutations in CYP21A2 gene are the cause of NC 21-OHD in these two patients. Nanopore sequencing technique is a reliable new detection method for patients with NC 21-OHD.
