Original Article
Genetic diagnosis of non-classical 21-hydroxylase deficiency by the new nanopore sequencing detection method
Yanjie Xia, Peng Dai, Huikun Duan, Panlai Shi, Shanshan Gao, Xueyu Guo, Ning Liu, Xiangdong Kong
Published 2023-01-19
Cite as Chin J Lab Med, 2023, 46(1): 74-80. DOI: 10.3760/cma.j.cn114452-20220727-00438
Abstract
ObjectiveTo 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.
MethodsClinical 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.
ResultsThe 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.
ConclusionsThe 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.
Key words:
Nanopore sequencing; 21-Hydroxylase deficiency; Congenital adrenal hyperplasia; CYP21A2 gene
Contributor Information
Yanjie Xia
Prenatal Diagnosis Center, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052,China
Peng Dai
Prenatal Diagnosis Center, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052,China
Huikun Duan
Prenatal Diagnosis Center, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052,China
Panlai Shi
Prenatal Diagnosis Center, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052,China
Shanshan Gao
Prenatal Diagnosis Center, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052,China
Xueyu Guo
Beijing Hope Group Biological technology CO.,LTD, Beijing 102200, China
Ning Liu
Prenatal Diagnosis Center, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052,China
Xiangdong Kong
Prenatal Diagnosis Center, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052,China
