NCJ Number
210771
Journal
Journal of Forensic Sciences Volume: 50 Issue: 4 Dated: July 2005 Pages: 809-825
Date Published
July 2005
Length
17 pages
Annotation
This report presents the results of a developmental validation study performed with the Quantifiler Human DNA Quantification Kit and the Quantifiler Y Human Male DNA Quantification Kit, following the guidelines of the Scientific Working Group on DNA Analysis Methods (SWGDAM).
Abstract
These two new commercial kits use "real-time" quantitative PCR (polymerase chain reaction) to detect and quantify human nuclear DNA in forensic samples. The kits are intended to address some of the limitations of older DNA quantification methods, which have limited accuracy and sensitivity and tend to be labor-intensive and time-consuming. The Quantifiler Human Kit assay detects total human genomic DNA (both male and female), and the Quantifiler Y Human Male Kit assay detects only male DNA. This report's description of the materials and methods used in the study addresses the sources and preparation of genomic DNA's used in the testing; DNA quantification using UV absorbance, dye intercalation, and slot blot hybridization methods; quantification kits' assays; PCR amplification kit genotyping assays; species specificity studies; sensitivity studies; the hematin model system for PCR inhibition; the preparation of degraded genomic DNA samples; the analysis of case type samples; DNA mixture studies; and accuracy and precision studies. The validation experiments showed the assays to be reliable and robust, producing quantification results consistent with other representative DNA quantification methods. The use of the Quantifiler kit assays as a preliminary procedure allowed a high success rate for subsequent sample analysis with a STR genotyping method. The assays can separately detect male DNA in mixed samples and provide more information about the presence of PCR inhibitors in samples. Also, the assays are frugal with samples. 7 figures, 5 tables and 27 references