Bite through the tent

The authors report on a young boy who was bitten into his face by an unknown animal while being asleep in a tent. Given the bite marks and the location of the scene, members of the mustelidae and canidae families were the first "suspects." Deoxyribunucleic acid (DNA) recovered from the tent's wall was analyzed with regard to parts of the mitochondrial 12S ribosomal ribunucleic acid (12S rRNA) and cytochrome b (cytb) genes as well as nuclear short tandem repeats (STRs). Since Sanger sequencing revealed a mixed sequence with a strong human component overlying the nonhuman contributor, an animal screening using a duplex real-time polymerase chain reaction (PCR) with an intercalating dye and melt curve analysis was employed. The results were later confirmed by cloning. The applied commercial canine STR kit verified the animal family (canidae) but did not help in discriminating the species due to cross-species amplification. In the presented case, the real-time PCR assay offered the cheapest and fastest method for animal family determination, which then allowed for an appropriate and sample-saving strategy to characterize the causative animal species.