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Technical strategy for monozygotic twin discrimination by single-nucleotide variants

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Abstract

Monozygotic (MZ) twins are theoretically genetically identical. Although they are revealed to accumulate mutations after the zygote splits, discriminating between twin genomes remains a formidable challenge in the field of forensic genetics. Single-nucleotide variants (SNVs) are responsible for a substantial portion of genetic variation, thus potentially serving as promising biomarkers for the identification of MZ twins. In this study, we sequenced the whole genome of a pair of female MZ twins when they were 27 and 33 years old to approximately 30 × coverage using peripheral blood on an Illumina NovaSeq 6000 Sequencing System. Potentially discordant SNVs supported by whole-genome sequencing were validated extensively by amplicon-based targeted deep sequencing and Sanger sequencing. In total, we found nine bona fide post-twinning SNVs, all of which were identified in the younger genomes and found in the older genomes. None of the SNVs occurred within coding exons, three of which were observed in introns, supported by whole-exome sequencing results. A double-blind test was employed, and the reliability of MZ twin discrimination by discordant SNVs was endorsed. All SNVs were successfully detected when input DNA amounts decreased to 0.25 ng, and reliable detection was limited to seven SNVs below 0.075 ng input. This comprehensive analysis confirms that SNVs could serve as cost-effective biomarkers for MZ twin discrimination.

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Data Availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Funding

This work was supported by grants from the National Natural Science Foundation of China (grant number 81871534) and the Ministry of Finance of China (grant number GY2021G-2).

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Author notes

  1. Weifen Sun Ziwei Wang, and Shubo Wen contributed equally to this work.

Authors and Affiliations

  1. Shanghai Key Laboratory of Forensic Medicine, Shanghai Forensic Service Platform, Academy of Forensic Science, Ministry of Justice, Shanghai, 200063, China

    Weifen Sun, Ziwei Wang, Shubo Wen, Ao Huang, Hui Li, Lei Jiang & Xiling Liu

  2. Shanghai Institute of Hematology, State Key Laboratory of Medical Genomics, National Research Center for Translational Medicine at Shanghai, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China

    Weifen Sun

  3. Department of Forensic Science, Medical School of Soochow University, Suzhou, 215123, China

    Ziwei Wang, Shubo Wen & Ao Huang

  4. National Center for Gene Research, State Key Laboratory of Plant Molecular Genetics, Center of Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai, 200233, China

    Qi Feng, Danlin Fan & Qilin Tian

  5. Department of Clinical Laboratory, Shanghai Children’s Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200062, China

    Dingding Han

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Contributions

Conceptualization, X. L.; methodology, Z. W., S. W., A. H., H. L., L. J., Q. F., D. F., and Q. T.; data curation, X. L., D. H., and W. S.; visualization, W. S.; writing—original draft preparation, W. S. and X. L.; writing—review and editing, W. S. and X. L.; funding acquisition, X. L. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Xiling Liu.

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Sun, W., Wang, Z., Wen, S. et al. Technical strategy for monozygotic twin discrimination by single-nucleotide variants. Int J Legal Med 138, 767–779 (2024). https://doi.org/10.1007/s00414-023-03150-7

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