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Developmental Validation of Feline, Bovine, Equine, and Cervid Quantitative PCR Assays *
Author(s) -
Lindquist Christina D.,
Evans Jeffery J.,
Wictum Elizabeth J.
Publication year - 2011
Publication title -
journal of forensic sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.715
H-Index - 96
eISSN - 1556-4029
pISSN - 0022-1198
DOI - 10.1111/j.1556-4029.2010.01605.x
Subject(s) - taqman , biology , genotyping , real time polymerase chain reaction , polymerase chain reaction , computational biology , dna , digital polymerase chain reaction , microbiology and biotechnology , gene , genetics , genotype
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.