Performance characteristics of chimerism testing by next generation sequencing.

Area of Science:

• Genetics
• Molecular Biology
• Forensic Science

Background:

• Chimerism testing analyzes biological sample mixtures, traditionally using short tandem repeat (STR) loci with capillary electrophoresis (CE).
• Next-generation sequencing (NGS) technology enables high-throughput analysis of sequence reads for more accessible mixture quantification.

Purpose of the Study:

• To evaluate a FORENSIC NGS platform for accurate chimerism percentage measurement.
• To compare NGS chimerism detection with traditional CE methods.
• To explore NGS utility in identifying solid organ transplant donors.

Main Methods:

• Utilized a FORENSIC NGS chimerism platform analyzing both STR and single nucleotide polymorphism (SNP) loci.
• Compared NGS results with established CE methods for chimerism quantification.
• Assessed accuracy through six proficiency testing surveys.

Main Results:

• Achieved accurate chimerism detection down to a 1% limit threshold using NGS.
• Demonstrated significant correlation between NGS and CE methods from 1% to 10% donor detection.
• Attained 100% accuracy in proficiency testing over six surveys.
• Successfully applied NGS for donor identification in solid organ transplantation using SNP profiles.

Conclusions:

• NGS provides a sensitive, reliable, and accurate alternative to CE for chimerism testing in clinical settings.
• The NGS platform offers expanded utility for donor selection in solid organ transplantation.
• NGS facilitates precise quantification of donor-recipient mixtures.