Presented at the 60th Annual Meeting of the American Academy of Forensic Sciences, February 18–23, 2008, in Washington, DC. Also presented in part at the 18th International Symposium on Human Identification, October 2–3, 2007, in Hollywood, CA.
Developmental Validation of Feline, Bovine, Equine, and Cervid Quantitative PCR Assays*
Article first published online: 11 NOV 2010
2010 American Academy of Forensic Sciences. Published 2010. This article is a U.S. Government work and is in the public domain in the U.S.A.
Journal of Forensic Sciences
Volume 56, Issue Supplement s1, pages S29–S35, January 2011
How to Cite
Lindquist, C. D., Evans, J. J. and Wictum, E. J. (2011), Developmental Validation of Feline, Bovine, Equine, and Cervid Quantitative PCR Assays. Journal of Forensic Sciences, 56: S29–S35. doi: 10.1111/j.1556-4029.2010.01605.x
- Issue published online: 3 JAN 2011
- Article first published online: 11 NOV 2010
- Received 21 Aug. 2009; and in revised form 21 Dec. 2009; accepted 30 Dec. 2009.
- forensic science;
- DNA quantification;
- real-time PCR;
- quantitative PCR;
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. Following the strategy employed for the canine qPCR assay, we developed individual assays to accurately quantify feline, bovine, equine, and cervid nuclear DNA. 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.