TY - JOUR
T1 - Quantitative real-time PCR (qPCR) assay for human-dog-cat species identification and nuclear DNA quantification
AU - Kanthaswamy, S.
AU - Premasuthan, A.
AU - Ng, J.
AU - Satkoski, J.
AU - Goyal, V.
N1 - Funding Information:
This study was supported by a National Institute of Justice grant 2008-DN-BX-K288 to SK. We wish to thank Joy Halverson (Questgen Forensics, Davis, CA), Eric Johnston (Scidera, Davis, CA) Marilyn Menotti (US National Cancer Research Institute), the UC Davis Veterinary Medicine Training Hospital (VMTH), and various human and non-human donors for their human, dog and cat samples. We would like to thank Vivek Goyal for his tremendous input in initiating the technical aspects of this project.
PY - 2012/3
Y1 - 2012/3
N2 - 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.
AB - 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.
KW - DNA quantification
KW - Human, dog and cat forensic DNA analysis
KW - Species identification
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U2 - 10.1016/j.fsigen.2011.06.005
DO - 10.1016/j.fsigen.2011.06.005
M3 - Article
C2 - 21764401
AN - SCOPUS:84857039953
SN - 1872-4973
VL - 6
SP - 290
EP - 295
JO - Forensic Science International: Genetics
JF - Forensic Science International: Genetics
IS - 2
ER -