Mitochondrial DNA-based identification of some forensically important blowflies in Thailand.
Accurate identification of insects collected from death scenes provides not only specific developmental data assisting forensic entomologists to determine the postmortem interval more precisely but also other kinds of forensic evidence. However, morphological identification can be complicated due to the similarity among species, especially in the early larval stages. To simplify and make the species identification more practical and reliable, DNA-based identification is preferentially considered. In this study, we demonstrate the application of partial mitochondrial cytochrome oxidase I (COI) and cytochrome oxidase II (COII) sequences for differentiation of forensically important blowflies in Thailand; Chrysomya megacephala, Chrysomya rufifacies and Lucilia cuprina by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The PCR yields a single 1324bp-sized amplicon in all blowfly specimens, followed by direct DNA sequencing. Taq(α)I and VspI predicted from the sequencing data provide different RFLP profiles among these three species. Sequence analysis reveals no significant intraspecific divergence in blowfly specimens captured from different geographical regions in Thailand. Accordingly, neighbor-joining tree using Kimura's 2-parameter model illustrates reciprocal monophyly between species. Thus, these approaches serve as promising tools for molecular identification of these three common forensically important blowfly species in Thailand.
Preativatanyou
Kanok
K
Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Rama IV Road, Bangkok 10330, Thailand.
Sirisup
Nantana
N
Payungporn
Sunchai
S
Poovorawan
Yong
Y
Thavara
Usavadee
U
Tawatsin
Apiwat
A
Sungpradit
Sivapong
S
Siriyasatien
Padet
P
D000818:Animals; D004272:DNA, Mitochondrial / Q000235:genetics*; D004175:Diptera / Q000235:genetics*; D003576:Electron Transport Complex IV / Q000235:genetics; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA; D013785:Thailand
D004272:DNA, Mitochondrial; D003576:Electron Transport Complex IV