Cost-Effective Next Generation Sequencing-Based STR Typing with Improved Analysis of Minor, Degraded and...

Area of Science:

• Forensic Science
• Genetics
• Molecular Biology

Background:

• Short tandem repeat (STR) analysis is crucial for forensic DNA profiling.
• Capillary electrophoresis (CE) is the standard method, but next-generation sequencing (NGS) offers advantages for complex samples.
• Current NGS forensic assays are often not cost-effective for low-throughput applications.

Purpose of the Study:

• To introduce and evaluate a cost-efficient shallow-sequence NGS assay (maSTR) for forensic STR typing.
• To compare the performance of the maSTR assay with a commercial CE-based kit.
• To assess the applicability of the maSTR assay in forensic and biomedical identification contexts.

Main Methods:

• Development of the maSTR assay for shallow-sequence NGS output.
• Implementation of the SNiPSTR bioinformatics pipeline for data analysis.
• Back-to-back comparison of maSTR with a commercial CE-based forensic STR kit using various sample types.

Main Results:

• The maSTR assay demonstrated comparable performance to CE for samples with low DNA content, mixed DNA, and PCR inhibitors.
• The maSTR assay showed superior performance compared to CE for degraded DNA samples.
• The maSTR assay, coupled with SNiPSTR, is a robust and cost-efficient NGS-based STR typing method.

Conclusions:

• The maSTR assay provides a viable, cost-effective NGS alternative for forensic STR profiling.
• This method enhances DNA analysis capabilities, particularly for challenging sample types.
• The maSTR assay is suitable for human identification in both forensic and biomedical fields.