Nanopore sequencing technology in forensic science: status quo and tendency

Next-generation sequencing (NGS) is widely used in forensics for high throughput but limited by short reads and high non-batch testing costs. Third-generation sequencing (TGS), especially Oxford Nanopore Technologies (ONT) nanopore sequencing, with ultra-long reads, portability, real-time sequencing and amplification-free detection, addresses NGS's forensic technical bottlenecks.

This study systematically reviews nanopore sequencing's current status in forensic genetics, explores its potential applications, clarifies its advantages and optimisation directions, and discusses practical challenges, to provide a comprehensive reference for its widespread forensic adoption.

Bibliometric analysis was conducted on English literature retrieved from PubMed, Web of Science and Scopus using keywords like "nanopore sequencing + forensic" (retrieval up to February 2026). Relevant literature was also summarised for technical principles, performance, applications and challenges.

Nanopore sequencing has unique forensic advantages (ultra-long reads for haplotype analysis, direct methylation/RNA detection, portability). Bibliometric analysis showed staged research development, with the Netherlands leading, Europe as the collaboration core, and hotspots shifting to frontier applications. Technical optimisations improved accuracy, but bottlenecks like low-input sample processing and standardisation remain.

Nanopore sequencing supplements traditional forensic sequencing with broad prospects. With hardware/bioinformatics optimisation, multi-laboratory validation and unified standards, it is expected to be widely used in routine forensics in 3-5 years, supporting judicial justice.