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A novel gross deletion and breakpoint junction sequence analysis of ATP7B in a Chinese family with Wilson disease using next‑generation sequencing and Sanger sequencing

  • Authors:
    • Wei‑Liang Liu
    • Fang Li
    • Lu Liu
    • Wei Chen
    • Zhi‑Xu He
    • Hao Gu
    • Rong Ai
  • View Affiliations / Copyright

    Affiliations: Department of Pediatrics, Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou 550004, P.R. China, Department of Ophthalmology, Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou 550004, P.R. China
    Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 517-523
    |
    Published online on: November 20, 2019
       https://doi.org/10.3892/mmr.2019.10830
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Abstract

Wilson disease (WD) is a rare autosomal recessive genetic disorder that causes abnormal copper metabolism, resulting in pathological accumulation of copper in the liver, brain and other organs. Mutations in the ATPase copper transporter 7B (ATP7B) gene, which encodes a membrane P‑type adenosine triphosphatase, have been identified as being responsible for WD. The present study analyzed clinical data and collected DNA samples from a pediatric patient with WD and her healthy parents. Mutation screening for ATP7B was performed using direct sequencing, multiplex ligation‑dependent probe amplification(MLPA), next‑generation sequencing (NGS) and Sanger sequencing of the breakpoint junction sequence. The patient (age, 2.7 years) presented with early‑onset hepatic disease. The present study identified compound heterozygous mutations of ATP7B, including a heterozygous mutation (p.Arg1,041Trp) and a novel heterozygous gross deletion of a 57,771 bp fragment (chr13: 52490972‑52548742) (GRCh37) from partial exon2‑ exon21 to external ATP7B sequence (15.833bp) in the patient. Analysis of the family members of the patient showed that the missense mutation and the gross deletion mutation were inherited from her mother and father, respectively. Microhomology and inverted repeat sequences, which may mediate the deletion mutation, were identified through sequence analysis on both sides of the breakpoints of this deletion. The present study provided additional information on the genotypic spectrum of the ATP7B gene, particularly with regard to early onset hepatic disease, as observed in the present patient with WD. The identification of the precise breakpoint junction sequence warrants further investigation of DNA break and recombination mechanisms. In detecting precise deletions, the NGS associated with Sanger sequencing of breakpoint junction sequence have been found to have more advantages than MLPA.
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Copy and paste a formatted citation
Spandidos Publications style
Liu WL, Li F, Liu L, Chen W, He ZX, Gu H and Ai R: A novel gross deletion and breakpoint junction sequence analysis of ATP7B in a Chinese family with Wilson disease using next‑generation sequencing and Sanger sequencing. Mol Med Rep 21: 517-523, 2020.
APA
Liu, W., Li, F., Liu, L., Chen, W., He, Z., Gu, H., & Ai, R. (2020). A novel gross deletion and breakpoint junction sequence analysis of ATP7B in a Chinese family with Wilson disease using next‑generation sequencing and Sanger sequencing. Molecular Medicine Reports, 21, 517-523. https://doi.org/10.3892/mmr.2019.10830
MLA
Liu, W., Li, F., Liu, L., Chen, W., He, Z., Gu, H., Ai, R."A novel gross deletion and breakpoint junction sequence analysis of ATP7B in a Chinese family with Wilson disease using next‑generation sequencing and Sanger sequencing". Molecular Medicine Reports 21.1 (2020): 517-523.
Chicago
Liu, W., Li, F., Liu, L., Chen, W., He, Z., Gu, H., Ai, R."A novel gross deletion and breakpoint junction sequence analysis of ATP7B in a Chinese family with Wilson disease using next‑generation sequencing and Sanger sequencing". Molecular Medicine Reports 21, no. 1 (2020): 517-523. https://doi.org/10.3892/mmr.2019.10830
Copy and paste a formatted citation
x
Spandidos Publications style
Liu WL, Li F, Liu L, Chen W, He ZX, Gu H and Ai R: A novel gross deletion and breakpoint junction sequence analysis of ATP7B in a Chinese family with Wilson disease using next‑generation sequencing and Sanger sequencing. Mol Med Rep 21: 517-523, 2020.
APA
Liu, W., Li, F., Liu, L., Chen, W., He, Z., Gu, H., & Ai, R. (2020). A novel gross deletion and breakpoint junction sequence analysis of ATP7B in a Chinese family with Wilson disease using next‑generation sequencing and Sanger sequencing. Molecular Medicine Reports, 21, 517-523. https://doi.org/10.3892/mmr.2019.10830
MLA
Liu, W., Li, F., Liu, L., Chen, W., He, Z., Gu, H., Ai, R."A novel gross deletion and breakpoint junction sequence analysis of ATP7B in a Chinese family with Wilson disease using next‑generation sequencing and Sanger sequencing". Molecular Medicine Reports 21.1 (2020): 517-523.
Chicago
Liu, W., Li, F., Liu, L., Chen, W., He, Z., Gu, H., Ai, R."A novel gross deletion and breakpoint junction sequence analysis of ATP7B in a Chinese family with Wilson disease using next‑generation sequencing and Sanger sequencing". Molecular Medicine Reports 21, no. 1 (2020): 517-523. https://doi.org/10.3892/mmr.2019.10830
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