title: Quantitative real-time PCR (qPCR) assay for human-dog-cat species identification and nuclear DNA quantification.
abstract: In the United States, human forensic evidence collected from crime scenes is usually comingled with biomaterial of canine and feline origins. Knowledge of the concentration of nuclear DNA extracted from a crime scene biological sample and the species from which the sample originated is essential for DNA profiling. The ability to accurately detect and quantify target DNA in mixed-species samples is crucial when target DNA may be overwhelmed by non-target DNA. We have designed and evaluated a species-specific (human, dog and cat) nuclear DNA identification assay based on the TaqMan(®) quantitative real-time PCR (qPCR) technology that can simultaneously detect and measure minute quantities of DNA specific to either humans, dogs and/or cats. The fluorogenic triplex assay employs primers and hydrolysis probes that target the human TH01 locus as well as the dog and cat Melanocortin 1 Receptor (MC1R) sequences in a species-specific manner. We also demonstrate that the assay is a highly sensitive, reliable and robust method for identifying and quantifying mixed-species templates of human-dog-cat origin with as little as 0.4 pg of human and cat nuclear DNA, respectively, and 4.0 pg of dog nuclear DNA.
authors.affiliation: Department of Environmental Toxicology, University of California Davis, Davis, CA 95616, USA. skanthaswamy@ucdavis.edu
authors.forename: S
authors.initials: S
authors.lastname: Kanthaswamy
authors.forename: A
authors.initials: A
authors.lastname: Premasuthan
authors.forename: J
authors.initials: J
authors.lastname: Ng
authors.forename: J
authors.initials: J
authors.lastname: Satkoski
authors.forename: V
authors.initials: V
authors.lastname: Goyal
mesh_terms: D000818:Animals; D002415:Cats / Q000235:genetics*; D002467:Cell Nucleus / Q000235:genetics; D004247:DNA / Q000235:genetics*; D016172:DNA Fingerprinting / Q000379:methods; D017931:DNA Primers; D015342:DNA Probes; D004285:Dogs / Q000235:genetics*; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction; D044102:Receptor, Melanocortin, Type 1 / Q000235:genetics; D015203:Reproducibility of Results; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D044102:Receptor, Melanocortin, Type 1; D004247:DNA