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Microhaplotypes in forensic genetics: From exploration to application in degraded DNA specimens
Chiara Turchi1, Filomena Melchionda1, Fabiano Gentile2
1Section of Legal Medicine, Department of Biomedical Sciences and Public Health, Polytechnic University of Marche, Via Tronto, Ancona, AN 60126, Italy.
Forensic Science International. Genetics
|November 21, 2025
Summary
This study developed a microhaplotype (MH) panel for forensic DNA analysis, proving effective for degraded samples, kinship identification, and ancestry inference with high sensitivity and accuracy.
Area of Science:
- Forensic genetics
- Population genetics
- Molecular biology
Background:
- Microhaplotypes (MHs) are increasingly vital forensic markers.
- Fragmented DNA in forensic samples poses analytical challenges.
Purpose of the Study:
- Develop a Massively Parallel Sequencing (MPS) panel of microhaplotypes for forensic applications.
- Evaluate the panel's efficacy in degraded DNA analysis, kinship identification, and ancestry inference.
Main Methods:
- Designed an MPS assay with amplicons <140 bp for degraded DNA.
- Established a 76-microhaplotype panel (299 SNPs).
- Genotyped 102 Italian individuals to determine allele and haplotype frequencies.
Main Results:
- The panel achieved complete profiles with as little as 0.05 ng DNA.
- High sensitivity was demonstrated on degraded forensic samples.
- Probabilistic genotyping yielded extremely strong likelihood ratios (7.84E+09 to 2.70E+34).
- Full- and half-sibling relationships were distinguishable.
- PCA and STRUCTURE analyses confirmed ancestry informative potential.
Conclusions:
- The developed microhaplotype panel is highly effective for forensic identification, even with degraded DNA.
- The panel offers significant power for kinship analysis and ancestry inference.
- Short microhaplotypes provide sensitive and reliable genetic profiles in challenging forensic casework.
