Use of Cytochrome c Oxidase Subunit I (COI) Nucleotide Sequences for Identification of the Korean Luciliinae Fly Species (Diptera: Calliphoridae) in Forensic Investigations

Abstract

Blowflies, especially species belonging to the subfamily Luciliinae, are the first insects to lay eggs on corpses in Korea. Fast and accurate species identification has been a key task for forensic entomologists. Because conventional morphologic identification methods have many limitations with respect to forensic practice, molecular methods have been proposed to identify fly species of forensic importance. To this end, the authors amplified and sequenced the full length of the cytochrome c oxidase subunit I (COI) gene of the Luciliinae fly species collected in Korea. The results showed the COI sequences are instrumental in identifying Luciliinae fly species. However, when compared with previously reported data, considerable inconsistencies were noted. Hemipyrellia ligurriens data in this study differed significantly from two of the five pre-existing data. Two closely related species, Lucilia illustris and Lucilia caesar, showed an overlap of COI haplotypes due to four European sequences. The results suggest that more individuals from various geographic regions and additive nuclear DNA markers should be analyzed, and morphologic identification keys must be reconfirmed to overcome these inconsistencies.

Using the Developmental GeneBicoidto Identify Species of Forensically Important Blowflies (Diptera: Calliphoridae)

“…The new 5′ primers and a 3′ primer (R2), which hybridized more than 200 bp downstream of the initial primer, were used for subsequent PCR amplifications [9, 10]. After analyzing the sequences from the first and second rounds of PCR, specific primers (FD1–9 and RD1–3) were designed (see supplementary Table 2) and used to amplify 500–525 bp targets.…”

“…Sequencing was performed with automatic sequencers (ABI PRISM 310 genetic analyzer) [9, 10]. Direct sequencing of amplified products was performed using the BigDye Terminator Sequencing kit (v1.1) (Applied Biosystems, Foster City, CA, USA).…”

Role of Polymerase Chain Reaction in Forensic Entomology

“…Subsequent to this research, mtDNA has been widely used for DNA analysis in forensic entomology, using COI and COII gene sequences analysis for distinguishing forensically important blow flies and flesh flies [4][5][6][7][8][9][10][11][12][13][14]. In purpose, such molecular work is similar to other non-entomological forensic methods in that it provides supplementary evidence in the form of PMI estimate to support the charge of a suspect to the crime.…”

Testing the utility of mitochondrial cytochrome oxidase subunit 1 sequences for phylogenetic estimates of relationships between crane (Grus) species

“…According to Funk and Omland (2003), at least 702 arthropod species are polyphyletic (26.5%), and 23% of 2319 animal species show species-level paraphyly or polyphyly in their mtDNA gene trees, including 16.7% of 331 bird species. Several explanations have been offered for this: the analyses may have consisted of inadequate sequence data or inaccurate taxonomy and/or may have included interspecific hybridisation or introgression, and incomplete sorting (Park et al, 2009). For example, the mtDNA inherited marker could be a result of hybridisation events, which results in morphology that differs from the mtDNA make-up of the individuall (Petersen et al, 2003).…”