title: Human DNA quantification and sample quality assessment: Developmental validation of the PowerQuant(®) system.
abstract: Quantification of the total amount of human DNA isolated from a forensic evidence item is crucial for DNA normalization prior to short tandem repeat (STR) DNA analysis and a federal quality assurance standard requirement. Previous commercial quantification methods determine the total human DNA and total human male DNA concentrations, but provide limited information about the condition of the DNA sample. The PowerQuant(®) System includes targets for quantification of total human and total human male DNA as well as targets for evaluating whether the human DNA is degraded and/or PCR inhibitors are present in the sample. A developmental validation of the PowerQuant(®) System was completed, following SWGDAM Validation Guidelines, to evaluate the assay's specificity, sensitivity, precision and accuracy, as well as the ability to detect degraded DNA or PCR inhibitors. In addition to the total human DNA and total human male DNA concentrations in a sample, data from the degradation target and internal PCR control (IPC) provide a forensic DNA analyst meaningful information about the quality of the isolated human DNA and the presence of PCR inhibitors in the sample that can be used to determine the most effective workflow and assist downstream interpretation.
authors.affiliation: Promega Corporation, 2800 Woods Hollow Rd., Madison, WI 53711, USA. Electronic address: margaret.ewing@promega.com.
authors.forename: Margaret M
authors.initials: MM
authors.lastname: Ewing
authors.affiliation: Promega Corporation, 2800 Woods Hollow Rd., Madison, WI 53711, USA. Electronic address: jonelle.thompson@promega.com.
authors.forename: Jonelle M
authors.initials: JM
authors.lastname: Thompson
authors.affiliation: Promega Corporation, 2800 Woods Hollow Rd., Madison, WI 53711, USA. Electronic address: bob.mclaren@promega.com.
authors.forename: Robert S
authors.initials: RS
authors.lastname: McLaren
authors.affiliation: Wisconsin State Crime Laboratory ∧ Madison, 4626 University Ave., Madison, WI 53705, USA. Electronic address: purperovm@doj.state.wi.us.
authors.forename: Vincent M
authors.initials: VM
authors.lastname: Purpero
authors.affiliation: Wisconsin State Crime Laboratory ∧ Madison, 4626 University Ave., Madison, WI 53705, USA. Electronic address: thomaskj@doj.state.wi.us.
authors.forename: Kelli J
authors.initials: KJ
authors.lastname: Thomas
authors.affiliation: Wisconsin State Crime Laboratory ∧ Milwaukee, 1578S 11th St., Milwaukee, WI 53204-2860, USA. Electronic address: dobrowskipa@doj.state.wi.us.
authors.forename: Patricia A
authors.initials: PA
authors.lastname: Dobrowski
authors.affiliation: Wisconsin State Crime Laboratory ∧ Milwaukee, 1578S 11th St., Milwaukee, WI 53204-2860, USA. Electronic address: degrootga@doj.state.wi.us.
authors.forename: Gretchen A
authors.initials: GA
authors.lastname: DeGroot
authors.affiliation: U.S. National Institute of Standards and Technology, 100 Bureau Drive, Gaithersburg, MD 20899-8314, USA. Electronic address: erica.romsos@nist.gov.
authors.forename: Erica L
authors.initials: EL
authors.lastname: Romsos
authors.affiliation: Promega Corporation, 2800 Woods Hollow Rd., Madison, WI 53711, USA. Electronic address: doug.storts@promega.com.
authors.forename: Douglas R
authors.initials: DR
authors.lastname: Storts
mesh_terms: D000818:Animals; D041322:Chromosomes, Human, Y; D004247:DNA / Q000032:analysis*; D053836:DNA Degradation, Necrotic; D016172:DNA Fingerprinting; D006801:Humans; D008297:Male; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D044469:Racial Groups / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D004247:DNA
keywords: DNA analysis; Forensic science; PowerPlex(®); PowerQuant(®); Quantification; Short tandem repeat (STR); Validation; qPCR