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Clinical validation and application of targeted long-range PCR and long-read sequencing-based analysis for haemophilia: experience from a haemophilia treatment centre in China.
Shi, Meizhen; Ma, Yunting; Peng, Xianwei; Zhou, Xu; Cheng, Zifeng; Xie, Bobo; Wei, Xianda; Gui, Chunrong; Mao, Aiping; Lin, Wenting; Luo, Jiefeng; Lai, Yinghui; Gui, Baoheng.
Afiliación
  • Shi M; Center for Medical Genetics and Genomics, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region, 530007, China; The Guangxi Health Commission Key Laboratory of Medical Genetics and Genomics, The Second Affiliated Hospital of Guangxi Medical Universit
  • Ma Y; The Second School of Medicine, Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region, 530007, China.
  • Peng X; Department of Hematology, The Second Affiliated Hospital of Guangxi Medical University.
  • Zhou X; The Second School of Medicine, Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region, 530007, China.
  • Cheng Z; The Second School of Medicine, Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region, 530007, China.
  • Xie B; Center for Medical Genetics and Genomics, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region, 530007, China; The Guangxi Health Commission Key Laboratory of Medical Genetics and Genomics, The Second Affiliated Hospital of Guangxi Medical Universit
  • Wei X; Center for Medical Genetics and Genomics, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region, 530007, China; The Guangxi Health Commission Key Laboratory of Medical Genetics and Genomics, The Second Affiliated Hospital of Guangxi Medical Universit
  • Gui C; Center for Medical Genetics and Genomics, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region, 530007, China; The Guangxi Health Commission Key Laboratory of Medical Genetics and Genomics, The Second Affiliated Hospital of Guangxi Medical Universit
  • Mao A; Berry Genomics Corporation, Beijing, 102200, China.
  • Lin W; Center for Medical Genetics and Genomics, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region, 530007, China; The Guangxi Health Commission Key Laboratory of Medical Genetics and Genomics, The Second Affiliated Hospital of Guangxi Medical Universit
  • Luo J; Department of Neurology, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region, 530007, China. Electronic address: drljf98@163.com.
  • Lai Y; Department of Hematology, The Second Affiliated Hospital of Guangxi Medical University. Electronic address: yinghuilai@sina.com.
  • Gui B; Center for Medical Genetics and Genomics, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region, 530007, China; The Guangxi Health Commission Key Laboratory of Medical Genetics and Genomics, The Second Affiliated Hospital of Guangxi Medical Universit
J Thromb Haemost ; 2024 Sep 09.
Article en En | MEDLINE | ID: mdl-39260745
ABSTRACT

BACKGROUND:

Targeted long-read sequencing (LRS) is expected to comprehensively analyse diverse complex variants in haemophilia A (HA) and B (HB), caused by the F8 and F9 genes, respectively. However, its clinical applicability still requires extensive validation.

OBJECTIVES:

To evaluate the clinical applicability of targeted LRS-based analysis, compared with routine PCR-based methods.

METHODS:

Gene variants of retrieved subjects were retrospectively and prospectively analysed. Whole-genome sequencing (WGS) was performed to further analyse undiagnosed cases. Breakpoints of novel genomic rearrangements were mapped and validated using long-distance-PCR and long-range-PCR combined with sequencing.

RESULTS:

Totally, 122 subjects were retrieved. In retrospective analysis of the 90 HA cases, HA-LRS assay showed consistent results in 84 cases compared with routine methods, and characterized six large deletions with their exact breakpoints confirmed by further validation in six cases (routine methods only presented failure in amplifying the involved exons). In prospective analysis of the 21 HA subjects, 20 variants of F8 were identified in 20 cases. For the remaining HA patient, no duplication/deletion or SNV/InDel was found, but a potential recombination involving exons 14 and 21 of F8 was observed by LRS. WGS analysis and further verification defined a 30,478bp tandem repeat involving exons 14-21 of F8. Among the 11 HB patients, HB-LRS analysis detected 11 SNVs/InDels in F9, consistent with routine methods.

CONCLUSIONS:

Targeted LRS-based analysis is efficient and comprehensive to identify SNVs/InDels and genomic rearrangements of haemophilia genes, especially we first expanding the panel including F9. However, further investigation for complex gross rearrangement is still essential.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: J Thromb Haemost Asunto de la revista: HEMATOLOGIA Año: 2024 Tipo del documento: Article Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: J Thromb Haemost Asunto de la revista: HEMATOLOGIA Año: 2024 Tipo del documento: Article Pais de publicación: Reino Unido