This technical report describes the internal validation process for the Applied Biosystems® AmpFlSTR® Yfiler™ PCR Amplification Kit, which is a multiplex assay that amplifies 17 loci located on the Y chromosome.
The authors of this report describe the internal validation process for the Applied Biosystems® AmpFlSTR® Yfiler™ PCR Amplification Kit, which is a multiplex assay that amplifies 17 loci located on the Y chromosome: DYS456, DYS389I, DYS390, DYS389II, DYS458, DYS19, DYS385a/b, DYS393, DYS391, DYS439, DYS635, DYS392, Y GATA H4, DYS437, DYS438, and DYS448. The authors describe the research design, materials and methods, and results and discussions. Eight studies were performed in this validation: analytical threshold; precision; contamination; sensitivity; reproducibility; concordance; mixtures; and stutter. For each study, results were derived from peak height relative fluorescence units (RFU) values and/or allele calls. The results demonstrated that the internal validation was successful and that the Yfiler™ kit successfully amplified evidence samples from adjudicated casework, is male specific, and is precise. The authors note that successful validation reinforces the manufacturer’s developmental validation performed prior to Yfiler™ being placed on the market, and that analysts will need to undergo proficiency testing before implementing the Yfiler™ kit into casework. The authors also recommend that a National Institute of Standards and Technology (NIST) Standard Reference Material (SRM) with a Y-STR profile, such as SRM 2395, should be analyzed to ensure quality assurance.
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