Abstract
Real-time PCR is most commonly used in the forensic community to quantify small amounts of human DNA in evidentiary samples. In the growing field of wildlife forensic genetics, real-time PCR is utilized primarily for identifying the species of origin from illegally traded animal by-products. Although DNA sequencing of mitochondrial DNA is the most common approach for species identification, it can be costly. Utilization of a multiplex real-time PCR assay can be a fast, inexpensive, and robust approach to species identification that can aid law enforcement in prosecuting crimes against animals. African (Loxodonta africana) and Asian (Elephas maximus) elephant populations are categorized under Appendix I or II of the Convention on the International Trade of Endangered Species (CITES), respectively. An Appendix I listing includes species that are threatened with extinction, thus trade of these plants and animals is highly restricted. Species not facing extinction that require extra attention and regulations so that they don’t become exploited and over utilized are listed in Appendix II. The primary reason for the decline of these two animals is the illegal trade of their ivory. Other reasons for the decline in the elephant population are deforestation and human conflict. In wildlife crime laboratories, oftentimes species of origin can be determined by morphology. This method is limited by the expertise of the taxonomist and the condition of the animal product. Ivory is commonly carved into small figurines and trinkets. Elephant meat, hair, and hide are traded which can make it difficult to identify the species. These limitations have led to the development of genetic tests to identify species of origin in wildlife investigations. African and Asian elephants do have highly similar genomes; however in portions of their cyt b gene, variation exists. In this study a dual-genus, real-time PCR assay to identify elephant DNA for forensic purposes was developed. Following the assay development, a rigorous developmental validation was conducted according to current community recommendations set forth by the Scientific Working Group for DNA Analysis and Methods (SWGDAM). The completion of this work provides an assay that can generate data of evidentiary quality for wildlife crime laboratories.