High clinical utility of long-read sequencing for precise diagnosis of congenital adrenal hyperplasia in 322 probands
Abstract Background The molecular genetic diagnosis of congenital adrenal hyperplasia (CAH) is very challenging due to the high homology between the CYP21A2 gene and its pseudogene CYP21A1P. Methodology This study aims to assess the clinical efficacy of targeted long-read sequencing (T-LRS) by compa...
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BMC
2025-01-01
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| Series: | Human Genomics |
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| Online Access: | https://doi.org/10.1186/s40246-024-00696-4 |
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| author | Yunpeng Wang Gaohui Zhu Danhua Li Yu Pan Rong Li Ting Zhou Aiping Mao Libao Chen Jing Zhu Min Zhu |
| author_facet | Yunpeng Wang Gaohui Zhu Danhua Li Yu Pan Rong Li Ting Zhou Aiping Mao Libao Chen Jing Zhu Min Zhu |
| author_sort | Yunpeng Wang |
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| description | Abstract Background The molecular genetic diagnosis of congenital adrenal hyperplasia (CAH) is very challenging due to the high homology between the CYP21A2 gene and its pseudogene CYP21A1P. Methodology This study aims to assess the clinical efficacy of targeted long-read sequencing (T-LRS) by comparing it with a control method based on the combined assay (NGS, Multiplex ligation-dependent probe amplification and Sanger sequencing) and to introduce T-LRS as a first-tier diagnostic test for suspected CAH patients to improve the precise diagnosis of CAH. Results A large cohort of 562 participants including 322 probands and 240 family members was enrolled for the perspective (96 probands) and prospective study (226 probands). The comparison analysis of T-LRS and control method have been performed. In the perspective study, 96 probands were identified using both the control method and T-LRS. Concordant results were detected in 85.42% (82/96) of probands. T-LRS performed more precise diagnosis in 14.58% (14/96) of probands. Among these, a novel 4141 kb deletion involving CYP21A2 and TNXB was established. A new diagnosis was improved by T-LRS. The duplications were also precisely identified to clarify the misdiagnosis by MLPA. In the prospective study, Variants were identified not only in CYP21A2 but also in HSD3B2 and CYP11B1 in 226 probands. Expand to 322 probands, the actual frequency of duplication haplotype (1.55%) could be calculated due to the accurate genotyping. Moreover, 75.47% of alleles with SNVs/indels, 22.20% of alleles with deletion chimeras. Conclusion T-LRS has higher resolution and reduced cost than control method with accurate diagnosis. The clinical utility of L-LRS could help to provide precision therapy to CAH patients, advance the life-long management of this complex disease and promote our understanding of CAH. |
| format | Article |
| id | doaj-art-75f7a3b1d83146bdb26e6fa6039556bd |
| institution | Kabale University |
| issn | 1479-7364 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Human Genomics |
| spelling | doaj-art-75f7a3b1d83146bdb26e6fa6039556bd2025-01-19T12:32:26ZengBMCHuman Genomics1479-73642025-01-0119111710.1186/s40246-024-00696-4High clinical utility of long-read sequencing for precise diagnosis of congenital adrenal hyperplasia in 322 probandsYunpeng Wang0Gaohui Zhu1Danhua Li2Yu Pan3Rong Li4Ting Zhou5Aiping Mao6Libao Chen7Jing Zhu8Min Zhu9Department of Endocrine and Metabolic Diseases, Children’s Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and DisordersDepartment of Endocrine and Metabolic Diseases, Children’s Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and DisordersBerry Genomics CorporationDepartment of Endocrine and Metabolic Diseases, Children’s Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and DisordersDepartment of Endocrine and Metabolic Diseases, Children’s Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and DisordersDepartment of Endocrine and Metabolic Diseases, Children’s Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and DisordersBerry Genomics CorporationBerry Genomics CorporationDepartment of Pediatric Research Institute, Children’s Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing Key Laboratory of PediatricsDepartment of Endocrine and Metabolic Diseases, Children’s Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and DisordersAbstract Background The molecular genetic diagnosis of congenital adrenal hyperplasia (CAH) is very challenging due to the high homology between the CYP21A2 gene and its pseudogene CYP21A1P. Methodology This study aims to assess the clinical efficacy of targeted long-read sequencing (T-LRS) by comparing it with a control method based on the combined assay (NGS, Multiplex ligation-dependent probe amplification and Sanger sequencing) and to introduce T-LRS as a first-tier diagnostic test for suspected CAH patients to improve the precise diagnosis of CAH. Results A large cohort of 562 participants including 322 probands and 240 family members was enrolled for the perspective (96 probands) and prospective study (226 probands). The comparison analysis of T-LRS and control method have been performed. In the perspective study, 96 probands were identified using both the control method and T-LRS. Concordant results were detected in 85.42% (82/96) of probands. T-LRS performed more precise diagnosis in 14.58% (14/96) of probands. Among these, a novel 4141 kb deletion involving CYP21A2 and TNXB was established. A new diagnosis was improved by T-LRS. The duplications were also precisely identified to clarify the misdiagnosis by MLPA. In the prospective study, Variants were identified not only in CYP21A2 but also in HSD3B2 and CYP11B1 in 226 probands. Expand to 322 probands, the actual frequency of duplication haplotype (1.55%) could be calculated due to the accurate genotyping. Moreover, 75.47% of alleles with SNVs/indels, 22.20% of alleles with deletion chimeras. Conclusion T-LRS has higher resolution and reduced cost than control method with accurate diagnosis. The clinical utility of L-LRS could help to provide precision therapy to CAH patients, advance the life-long management of this complex disease and promote our understanding of CAH.https://doi.org/10.1186/s40246-024-00696-4Congenital adrenal hyperplasiaLong-read sequencingComparative analysisClinical utilityPrecise diagnosis |
| spellingShingle | Yunpeng Wang Gaohui Zhu Danhua Li Yu Pan Rong Li Ting Zhou Aiping Mao Libao Chen Jing Zhu Min Zhu High clinical utility of long-read sequencing for precise diagnosis of congenital adrenal hyperplasia in 322 probands Human Genomics Congenital adrenal hyperplasia Long-read sequencing Comparative analysis Clinical utility Precise diagnosis |
| title | High clinical utility of long-read sequencing for precise diagnosis of congenital adrenal hyperplasia in 322 probands |
| title_full | High clinical utility of long-read sequencing for precise diagnosis of congenital adrenal hyperplasia in 322 probands |
| title_fullStr | High clinical utility of long-read sequencing for precise diagnosis of congenital adrenal hyperplasia in 322 probands |
| title_full_unstemmed | High clinical utility of long-read sequencing for precise diagnosis of congenital adrenal hyperplasia in 322 probands |
| title_short | High clinical utility of long-read sequencing for precise diagnosis of congenital adrenal hyperplasia in 322 probands |
| title_sort | high clinical utility of long read sequencing for precise diagnosis of congenital adrenal hyperplasia in 322 probands |
| topic | Congenital adrenal hyperplasia Long-read sequencing Comparative analysis Clinical utility Precise diagnosis |
| url | https://doi.org/10.1186/s40246-024-00696-4 |
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