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Article

A multiplex ligation‑dependent probe amplification‑based next‑generation sequencing approach for the detection of copy number variations in the human genome

  • Authors:
    • Yongchen Yang
    • Chaoran Xia
    • Zaiwei Zhou
    • Dongkai Wei
    • Kangping Xu
    • Jia Jia
    • Wuhen Xu
    • Hong Zhang
  • View Affiliations / Copyright

    Affiliations: Department of Laboratory Medicine, Children's Hospital of Shanghai, Shanghai Jiao Tong University, Shanghai 200040, P.R. China, Shanghai Institute of Medical Genetics, Children's Hospital of Shanghai, Shanghai Jiao Tong University, Shanghai 200040, P.R. China, Product Department, WuXi Health Net Co., Ltd., Shanghai 200131, P.R. China, BasePair Biotechnology Co., Ltd., Suzhou, Jiangsu 215028, P.R. China, Shanghai Center for Bioinformation Technology, Shanghai Institutes of Biomedicine, Shanghai Academy of Science and Technology, Shanghai 201203, P.R. China
  • Pages: 5823-5833
    |
    Published online on: October 24, 2018
       https://doi.org/10.3892/mmr.2018.9581
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Abstract

The aim of the present study was to describe a multiplex ligation‑dependent probe amplification (MLPA)‑based next‑generation sequencing (NGS) assay that exhibited a significantly higher efficiency in detecting copy number variations (CNVs) and known single‑nucleotide variants, compared with traditional MLPA. MLPA polymerase chain reaction products were used to construct a library with indexed adapters, which was subsequently tested on an NGS platform, and the resulting data were analyzed by a series of analytical software. The reads from each probe reflected genetic variations in the target regions, and fragment differentiation was based on the specific base composition of the sequences, rather than fragment length, which was determined by capillary electrophoresis. The results of this approach were not only consistent with the MLPA results following capillary electrophoresis, but also coincided with the CNV results from the single‑nucleotide polymorphism array chip. This method allowed high‑throughput screening for the number of fragments and samples by integrating additional indices for detection. Furthermore, this technology precisely and accurately performed large‑scale detection and quantification of DNA variations, thereby serving as an effective and sensitive method for diagnosing genetic disorders caused by CNVs and known single‑nucleotide variations. Notably, MLPA‑NGS circumvents the problems associated with the inaccuracies of NGS in CNV detection due to the use of target sequence capture.
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Copy and paste a formatted citation
Spandidos Publications style
Yang Y, Xia C, Zhou Z, Wei D, Xu K, Jia J, Xu W and Zhang H: A multiplex ligation‑dependent probe amplification‑based next‑generation sequencing approach for the detection of copy number variations in the human genome. Mol Med Rep 18: 5823-5833, 2018.
APA
Yang, Y., Xia, C., Zhou, Z., Wei, D., Xu, K., Jia, J. ... Zhang, H. (2018). A multiplex ligation‑dependent probe amplification‑based next‑generation sequencing approach for the detection of copy number variations in the human genome. Molecular Medicine Reports, 18, 5823-5833. https://doi.org/10.3892/mmr.2018.9581
MLA
Yang, Y., Xia, C., Zhou, Z., Wei, D., Xu, K., Jia, J., Xu, W., Zhang, H."A multiplex ligation‑dependent probe amplification‑based next‑generation sequencing approach for the detection of copy number variations in the human genome". Molecular Medicine Reports 18.6 (2018): 5823-5833.
Chicago
Yang, Y., Xia, C., Zhou, Z., Wei, D., Xu, K., Jia, J., Xu, W., Zhang, H."A multiplex ligation‑dependent probe amplification‑based next‑generation sequencing approach for the detection of copy number variations in the human genome". Molecular Medicine Reports 18, no. 6 (2018): 5823-5833. https://doi.org/10.3892/mmr.2018.9581
Copy and paste a formatted citation
x
Spandidos Publications style
Yang Y, Xia C, Zhou Z, Wei D, Xu K, Jia J, Xu W and Zhang H: A multiplex ligation‑dependent probe amplification‑based next‑generation sequencing approach for the detection of copy number variations in the human genome. Mol Med Rep 18: 5823-5833, 2018.
APA
Yang, Y., Xia, C., Zhou, Z., Wei, D., Xu, K., Jia, J. ... Zhang, H. (2018). A multiplex ligation‑dependent probe amplification‑based next‑generation sequencing approach for the detection of copy number variations in the human genome. Molecular Medicine Reports, 18, 5823-5833. https://doi.org/10.3892/mmr.2018.9581
MLA
Yang, Y., Xia, C., Zhou, Z., Wei, D., Xu, K., Jia, J., Xu, W., Zhang, H."A multiplex ligation‑dependent probe amplification‑based next‑generation sequencing approach for the detection of copy number variations in the human genome". Molecular Medicine Reports 18.6 (2018): 5823-5833.
Chicago
Yang, Y., Xia, C., Zhou, Z., Wei, D., Xu, K., Jia, J., Xu, W., Zhang, H."A multiplex ligation‑dependent probe amplification‑based next‑generation sequencing approach for the detection of copy number variations in the human genome". Molecular Medicine Reports 18, no. 6 (2018): 5823-5833. https://doi.org/10.3892/mmr.2018.9581
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