Development and validation of a forensic multiplex InDel assay: The AGCU InDel 60 kit

Area of Science:

• Forensic Science
• Genetics
• Molecular Biology

Background:

• Insertion/deletion (InDel) polymorphisms are valuable genetic markers for human identification in forensics.
• Existing InDel kits offer complementary or stand-alone capabilities alongside short tandem repeats (STRs).

Purpose of the Study:

• To develop and validate a novel multiplex amplification detection system, the AGCU InDel 60 kit, for enhanced human identification.
• To evaluate the kit's performance in terms of accuracy, sensitivity, specificity, reproducibility, and robustness.

Main Methods:

• The AGCU InDel 60 kit was developed, containing 57 autosomal InDels, 2 Y-chromosomal InDels, and the amelogenin locus.
• Validation involved rigorous testing of PCR conditions, sensitivity, species specificity, reproducibility, stability, mock case samples, degradation, and population studies.
• A population study analyzed 564 unrelated individuals from three Han groups.

Main Results:

• The AGCU InDel 60 kit demonstrated accuracy, specificity, reproducibility, stability, and robustness.
• Complete DNA profiles were achieved with as little as 125 pg of human DNA.
• The kit detected significantly more alleles in degraded samples compared to an STR-based kit.
• Combined power of discrimination and exclusion values were exceptionally high, exceeding previous InDel kits.

Conclusions:

• The AGCU InDel 60 kit is a highly efficient and powerful tool for human identification in forensic applications.
• Its superior performance with degraded DNA and high discrimination power make it a valuable asset.
• The kit also serves as a supplementary tool for population genetics research.