Casework application of a stand-alone pentaplex assay of extended-ESS STRs

Using a stand-alone pentaplex comprising two standard-length short tandem repeats (STRs): D12S391 and D1S1656 plus three mini-STRs: D2S441, D10S1248 and D22S1045, all recently adopted to extend the European Standard Set (ESS) STRs, we have examined the genotyping performance of the new markers in 111 challenging casework samples. Although commercial kits now combine the five new STRs with existing core loci, we found the ESS-pentaplex we developed in-house performed better than both MiniFiler (comprising eight miniaturized STRs) and the NGM kit that includes the new STRs in a 15-marker multiplex. Our findings suggest at least part of the improved sensitivity of recently available ESS STRs can be attributed to the loci themselves as well as applying long-standing, robust primer designs that were first designed for the extended ESS markers by the laboratories that originally developed them. Therefore the ESS-pentaplex provides an ideal adjunct to Identifiler or MiniFiler to allow laboratories to assess the new STRs alongside existing standard loci, measure performance with challenging material and generate population frequency data ahead of a final decision on which additional STRs will extend the reconfigured CODIS core set.