Developmental validation of Applied Biosystems YFiler Platinum Casework PCR Amplification Kit

Abstract

Abstract The YFiler Platinum Casework PCR Amplification Kit is a 6-dye multiplex assay that simultaneously amplifies a set of 38 male-specific, Y-chromosome Short Tandem Repeat (YSTR) markers (DYS576, DYS389I, DYS635, DYS389II, DYS627, DYS549, DYS593, DYS645, DYS460, DYS458, DYS19, YGATAH4, DYS448, DYS391, DYS557, DYS522, DYS456, DYS390, DYS438, DYS392, DYS518, DYS444, DYS533, DYS570, DYS437, DYS385, DYS449, DYS643, DYS596, DYS393, DYS439, DYS481, DYF387S1, DYS527, DYS447), three insertion/deletion polymorphic markers (Yindels: rs771783753, rs759551978, rs199815934), and an internal quality control (IQC) system. When compared to the YFiler Platinum PCR Amplification kit for database samples, YFiler Platinum Casework kit was developed to include an improved Primer Mix incorporating a brighter TED dye, an updated internal quality control system, better resolution of large DNA fragments in Applied Biosystems POP-4 Polymer, and reduced female DNA cross-reactivity. Here, we report the results of the developmental validation study which followed the Scientific Working Group on DNA Analysis Methods (SWGDAM) guidelines and includes data for PCR-based studies, sensitivity, species specificity, stability, precision, reproducibility and repeatability, population concordance, stutter, DNA mixtures, and performance on mock casework samples. The results validate the multiplex design as well as demonstrate the kit’s robustness, reliability, and suitability as an assay for human identification with casework DNA samples.