Validation of a DNA-based method for identifying Chrysomyinae (Diptera : Calliphoridae) used in a death investigation

Many authors have proposed DNA-based methods for identifying an insect specimen associated with human remains. However, almost no attempt has been made to validate these methods using additional observations. We tested a protocol for identifying insects in the blow fly subfamily Chrysomyinae (Diptera: Calliphoridae) often found to be associated with a human corpse in Canada or the USA. This method uses phylogenetic analysis of DNA sequence from a short segment of the mitochondrial gene for cytochrome oxidase one (COI). Test chrysomyine COI sequences were obtained from 245 newly sequenced specimens and 51 specimens from the published literature. Published sequences from representatives of nonchrysomyine genera were also included to check for the possibility of a false positive identification. All of the chrysomyine test haplotypes were correctly identified with strong statistical support, and there were no false positives. This method appears to be an accurate and robust technique for identifying chrysomyine species from a death investigation in this geographic region. The far northern species Protophormia atriceps was not evaluated; therefore, caution is required in applying this method at very high latitudes in North America.