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Peer-reviewedCC BYGenes

Research Progress on the Application of Nanopore Long-Read Sequencing Combined with DNA Methylation Marking in Complex Forensic Evidence.

  • Qiannan Xu1,2,3
  • Xuanman Cui1,2,3
  • Shuyan Mei1,2,3
  • Haojie Qin1,2,3
  • Zhe Zheng1,2,3
  • Xiandun Zhai1,2,3
  1. 1School of Basic Medicine and Forensic Medicine, Henan University of Science and Technology, Luoyang 471023, China.
  2. 2Institute of Medical Aspects of Specific Environments, Henan University of Science and Technology, Luoyang 471023, China.
  3. 3Forensic Medicine Identification Center, Henan University of Science and Technology, Luoyang 471023, China.
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Abstract

Background In the analysis of complex forensic material evidence, conventional detection methods for highly degraded, trace, or mixed samples present several limitations, such as low typing success rates and limited information dimensions. Methods This study systematically examines the technical principles and compatibility of nanopore long-read sequencing combined with DNA methylation profiling, identifies optimization strategies to enhance sensitivity for trace materials, improves accuracy in methylation site interpretation, for instance, NanoFreeLunch, without raw signals and using base quality/error patterns, reaching 0.97-0.99 correlation with benchmark regional methylation levels, and streamlines library preparation workflows while evaluating its practical efficacy through domestic and international case studies. Results The research demonstrates that this technology enables simultaneous acquisition of genomic sequences and epigenetic information, facilitating multidimensional identification-including personal identification, age estimation, and body fluid identification-even in highly degraded samples, with significantly higher site detection rates compared to traditional methods. Conclusions Future efforts should focus on standardizing the technology and developing specialized databases to further refine its application frameworks for forensic investigations, thereby providing robust technical support for complex forensic evidence analysis.

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Cite

BibTeXRIS
BibTeX
@article{xu2026research,
  title = {{Research Progress on the Application of Nanopore Long-Read Sequencing Combined with DNA Methylation Marking in Complex Forensic Evidence.}},
  author = {Qiannan Xu and Xuanman Cui and Shuyan Mei and Haojie Qin and Zhe Zheng and Xiandun Zhai},
  journal = {Genes},
  year = {2026},
  month = oct,
  doi = {10.3390/genes17101235},
  url = {https://europepmc.org/article/MED/42938351},
  note = {CC BY},
}
RIS
TY  - JOUR
TI  - Research Progress on the Application of Nanopore Long-Read Sequencing Combined with DNA Methylation Marking in Complex Forensic Evidence.
AU  - Qiannan Xu
AU  - Xuanman Cui
AU  - Shuyan Mei
AU  - Haojie Qin
AU  - Zhe Zheng
AU  - Xiandun Zhai
PY  - 2026
DA  - 2026/10/07
JO  - Genes
DO  - 10.3390/genes17101235
UR  - https://europepmc.org/article/MED/42938351
AB  - Background In the analysis of complex forensic material evidence, conventional detection methods for highly degraded, trace, or mixed samples present several limitations, such as low typing success rates and limited information dimensions. Methods This study systematically examines the technical principles and compatibility of nanopore long-read sequencing combined with DNA methylation profiling, identifies optimization strategies to enhance sensitivity for trace materials, improves accuracy in methylation site interpretation, for instance, NanoFreeLunch, without raw signals and using base quality/error patterns, reaching 0.97-0.99 correlation with benchmark regional methylation levels, and streamlines library preparation workflows while evaluating its practical efficacy through domestic and international case studies. Results The research demonstrates that this technology enables simultaneous acquisition of genomic sequences and epigenetic information, facilitating multidimensional identification-including personal identification, age estimation, and body fluid identification-even in highly degraded samples, with significantly higher site detection rates compared to traditional methods. Conclusions Future efforts should focus on standardizing the technology and developing specialized databases to further refine its application frameworks for forensic investigations, thereby providing robust technical support for complex forensic evidence analysis.
SN  - 2073-4425
ER  -