Open Access

Non‑invasive prenatal diagnosis of thalassemia through multiplex PCR, target capture and next‑generation sequencing

  • Authors:
    • Xu Yang
    • Yanchou Ye
    • Dongmei Fan
    • Sheng Lin
    • Ming Li
    • Hongying Hou
    • Jun Zhang
    • Xuexi Yang
  • View Affiliations

  • Published online on: June 15, 2020     https://doi.org/10.3892/mmr.2020.11234
  • Pages: 1547-1557
  • Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Prenatal clinical detection of thalassemia involves gap‑PCR and reverse dot blot (RDB) analysis of fetal DNA acquired through invasive methods. The present study aimed to develop a non‑invasive prenatal diagnostic method for thalassemia based on next‑generation sequencing (NGS). A total of eight families with proband children with thalassemia were recruited for the study during a subsequent pregnancy. The sequence of the thalassemia genes of the parents and proband were determined using NGS, based on a thalassemia AmpliSeq panel. Cell‑free plasma DNA from pregnant women related to the aforementioned proband was analyzed using an NGS panel, based on thalassemia‑associated capture probes. Heterozygous single nucleotide polymorphisms within the 10 kb regions flanking exons of the targeted thalassemia genes were acquired using probes or AmpliSeq and employed for parental haplotype construction using Trio‑based panel sequencing. The fetal haplotype was deduced from the parental haplotypes and relative haplotype dosage, and subsequently validated using gap‑PCR and RDB, based on invasively sampled amniotic fluid. A non‑invasive prenatal diagnosis procedure from maternal plasma fetal DNA was successfully developed based on haplotype analysis. The deduced haplotypes of eight fetuses were identical to the results of invasive prenatal diagnosis procedures, with an accuracy rate of 100%. Taken together, the present study demonstrated the potential for non‑invasive prenatal diagnosis of α‑ and β‑thalassemia using NGS and haplotype‑assisted analysis.

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August-2020
Volume 22 Issue 2

Print ISSN: 1791-2997
Online ISSN:1791-3004

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Spandidos
Yang X, Ye Y, Fan D, Lin S, Li M, Hou H, et al: Non‑invasive prenatal diagnosis of thalassemia through multiplex PCR, target capture and next‑generation sequencing. Mol Med Rep 22: 1547-1557, 2020
APA
Yang, X., Ye, Y., Fan, D., Lin, S., Li, M., Hou, H. ... Yang, X. (2020). Non‑invasive prenatal diagnosis of thalassemia through multiplex PCR, target capture and next‑generation sequencing. Molecular Medicine Reports, 22, 1547-1557. https://doi.org/10.3892/mmr.2020.11234
MLA
Yang, X., Ye, Y., Fan, D., Lin, S., Li, M., Hou, H., Zhang, J., Yang, X."Non‑invasive prenatal diagnosis of thalassemia through multiplex PCR, target capture and next‑generation sequencing". Molecular Medicine Reports 22.2 (2020): 1547-1557.
Chicago
Yang, X., Ye, Y., Fan, D., Lin, S., Li, M., Hou, H., Zhang, J., Yang, X."Non‑invasive prenatal diagnosis of thalassemia through multiplex PCR, target capture and next‑generation sequencing". Molecular Medicine Reports 22, no. 2 (2020): 1547-1557. https://doi.org/10.3892/mmr.2020.11234