Developmental Validation of Feline, Bovine, Equine, and Cervid Quantitative PCR Assays

• NCJ Number: 233492
• Journal: Journal of Forensic Sciences Volume: 56 Issue: S1 Dated: January 2011 Pages: S29-S35
• Author(s): Christina D. Lindquist, M.Sc.; Jeffery J. Evans, B.Sc.; Elizabeth J. Wictum, B.Sc.
• Date Published: January 2011
• Length: 7 pages
• Publication Format: Article
• Publication Type: Report (Study/Research)
• Language: English
• Country: United States of America

Annotation

Following the strategy employed for the canine qPCR assay, this study developed individual assays to accurately quantify feline, bovine, equine, and cervid nuclear DNA.

Abstract

Accurate DNA quantification is essential for optimizing DNA testing and minimizing sample consumption. Real-time quantitative polymerase chain reaction (qPCR) assays have been published for human and canine nuclear DNA, and the need for quantifying other forensically important species was evident. The current study found that each TaqMan-based assay incorporates a genus-specific probe targeting the Melanocortin-1 Receptor gene and includes a piece of synthetic DNA that acts as an internal PCR control for detecting inhibition. Developmental validations were carried out following the revised guidelines of the Scientific Working Group on DNA Analysis Methods with modifications necessary for validation of nonhuman qPCR assays. All assays demonstrated the specificity, sensitivity, stability, reproducibility, accuracy, and precision required for forensic casework. The application of these assays to animal forensic DNA analysis has both conserved laboratory resources and improved genotyping results. (Published Abstract)