Retrofitting massively parallel sequencing (MPS) for HLA-DQA1 and polymarker (PM) in forensic casework

Area of Science:

• Forensic Science
• Genetics
• Molecular Biology

Background:

• Genotype profiling is crucial in forensics, with technology evolving from serology to DNA sequencing.
• Older forensic DNA typing methods, like allele-specific oligonucleotide hybridization assays from the 1990s, may require re-analysis with modern techniques.

Purpose of the Study:

• To develop a novel multiplex system combining PCR and massively parallel sequencing (MPS) for re-analyzing historical forensic DNA data.
• To address and resolve ambiguities in genotype assignment caused by paralogous genes in older forensic assays.

Main Methods:

• Development of a multiplex PCR system.
• Application of massively parallel sequencing (MPS) for DNA polymorphism identification.
• Implementation of strategies to overcome genotype assignment ambiguities from paralogous genes (GYPA and HBG2).

Main Results:

• The developed multiplex system successfully identified DNA polymorphisms.
• Results were consistent with the widely used AmpliType PM+DQA1 Amplification and Typing Kit.
• Technical solutions were devised to manage confounding factors from paralogous genes, ensuring accurate genotype assignment.

Conclusions:

• A novel resource is established for re-analyzing historical forensic DNA profiles using current technology.
• The study enhances the reliability of forensic DNA data interpretation, particularly for older samples.
• This advancement supports the re-evaluation of past forensic investigations with improved accuracy.