Quantitative real-time PCR (qPCR) assay for human-dog-cat species identification and nuclear DNA quantification

S. Kanthaswamy, A. Premasuthan, J. Ng, J. Satkoski, V. Goyal

Research output: Contribution to journalArticle

22 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)290-295
Number of pages6
JournalForensic Science International: Genetics
Volume6
Issue number2
DOIs
StatePublished - Mar 1 2012
Externally publishedYes

Keywords

  • DNA quantification
  • Human, dog and cat forensic DNA analysis
  • Species identification

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Genetics

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