Performance Characteristics of Commercial Y‐STR Multiplex Systems*

Abstract:  In this work, a number of performance checks were carried out to evaluate the efficacy of commercial Y‐short tandem repeats (Y‐STR) kits for casework applications. The study evaluated the sensitivity, specificity and stability of the Y‐STR markers used and the ability to obtain a male profile from postcoital samples taken at various time points after intercourse. All systems performed well with 1–3 ng of male DNA as recommended by the manufacturers. All systems gave full profiles at 100 pg of input DNA, which is within the realm of low copy number DNA analysis. Moreover all, except Y‐Plex™ 12, gave full profiles with 30–50 pg of male DNA. No increased performance was obtained with any of the systems by increasing the cycle number beyond that recommended by the various manufacturers. When up to 1 μg of female DNA was used (in the absence of male DNA) no female DNA cross reactivity was observed with the Y‐Plex™ 12 and Y‐Filer™ systems. PowerPlex® Y produced female DNA derived products near the DYS438 and within the DYS392 loci at a rare allele position with high input DNA levels (300 ng and 1 μg, respectively). Male/female DNA admixture experiments indicated the particularly high specificity of the Y‐Filer™ and PowerPlex® Y systems under conditions of several thousand fold female DNA excess. All systems were able to detect the minor alleles in male/male DNA admixtures at a 1:5 dilution with the PowerPlex® Y and Y‐Filer™ being able to detect some minor alleles at 1:20. Species testing indicated some limited, minor cross reactivity of the commercial systems with some domestic male mammals although it is easily recognizable and would not pose any problems in casework analysis. As expected a significant number of cross‐reacting products were obtained with nonhuman primate species. All Y‐STR multiplex systems tested were able to produce complete Y‐STR profiles from bloodstains and semen stains exposed up to 6 weeks when the samples were protected against precipitation and sunlight. However, exposure of the samples to precipitation either in the presence or absence of sunlight resulted in Y‐STR profile loss over time, with total profile loss occurring with all systems after 3 weeks or more. Complete Y‐STR profiles of the male donors up to 72 h postcoitus were obtained with all of the multiplex systems tested, except for Y‐Plex™ 12, which gave partial profiles.