Developmental Validation of the PowerPlex 16 HS System: An Improved 16-Locus Fluorescent STR Multiplex

• NCJ Number: 231569
• Journal: Forensic Science International: Genetics Volume: 4 Issue: 4 Dated: July 2010 Pages: 257-264
• Author(s): Martin G. Ensenberger; Jonelle Thompson; Becky Hill; Kristen Homick; Veronica Kearney; Kathleen A. Mayntz-Press; Paul Mazur; Amy McGuckian; Jelena Myers; Kelli Raley; Stewart G. Raley; Robin Rothove; Jonathan Wilson; Doug Wieczorek; Patricia M. Fulmer; Douglas R. Storts; Benjamin E. Krenke
• Date Published: July 2010
• Length: 8 pages
• Publication Format: Article
• Publication Type: Report (Study/Research)
• Language: English
• Country: Ireland

Annotation

This paper presents data from eight laboratories, including developmental data from Promega Corporation, that show the capabilities and limitations of the PowerPlex 16 HS System, an updated version of the PowerPlex 16 System, which first demonstrated single-reaction analysis of the 13 core loci for CODIS, the U.S. national DNA database system.

Abstract

The primers and dyes remain the same in both the original and updated systems; however, the PowerPlex 16 HS System introduces an enhanced buffer system that includes hot-start Taq DNA polymerase, ensuring robust performance. Due to the modification of the reaction mix, a multi-laboratory developmental validation study was conducted in order to document performance capabilities and limitations for the revised assay. The validation study found that genotyping of single-source samples was consistent across a large range of template DNA concentrations, with most laboratories obtaining complete profiles at 62.5 pg. Mixture analyses showed that over 90 percent of minor alleles were detected at 1:9 ratios. Optimum amplification cycle number was ultimately dependent on the sensitivity of the detection instrument and could be adjusted to accommodate a range of DNA template concentrations. Reaction conditions - including volume and annealing temperature, the concentrations of primers, Taq DNA polymerase, and magnesium - were optimal and able to withstand moderate variations without affecting multiplexed STR amplification. Data from nonprobative samples and concordance studies showed consistent results when comparing the PowerPlex HS System with the PowerPlex 16 System, along with other commercially available systems. This validation study, which was conducted according to SWGDAM guidelines, supports the use of the PowerPlex 16 HS System for forensic use. Materials and methods used are described. 10 figures and 27 references