Molecular Species Identification of Forensically Important Flies in Korea

Seong Hwan Park; Sang Eon Shin

doi:10.7580/kjlm.2013.37.4.177

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Park and Shin: Molecular Species Identification of Forensically Important Flies in Korea

종 설

Korean J Leg Med 2013;37(4):177-182.

Published online: 17 January 2013

DOI: https://doi.org/10.7580/kjlm.2013.37.4.177

Molecular Species Identification of Forensically Important Flies in Korea

Seong Hwan Park, Sang Eon Shin

Department of Legal Medicine, Korea University College of Medicine, Seoul, Korea

책임저자 : 박성환 (136-705) 서울특별시 성북구 인촌로 73 고려대학교 의과대학 법의학교실 전화 : +82-2-2286-1158 FAX : +82-2-928-3901 E-mail : kuforen@gmail.com

Received 25 October 2013 Accepted 11 November 2013

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

To estimate the postmortem interval (PMI) by using entomological evidence, species identification of forensically important flies is mandatory. However, the traditional species identification method, which relies on the key morphological features of adult flies, is not always available to investigators and has limitations to the immature samples. Because of these limitations, species identification using DNA sequences has long been an issue in the field of forensic entomology. In this review, I have briefly described the basic principles of molecular species identification and phylogenetic analysis and their applications in forensic entomology. I also recommend an experimental and statistical method to identify unknown fly samples obtained from the field.

Keywords: Key Words, Forensic science, Entomology, Species specificity, Phylogenetic DNA barcode

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Fig. 1.

An example of a neighbor-joining phylogenetic tree, using COI sequences of modern human (Homo sapiens), Neanderthal man (Homo neanderthalensis), chimpanzee (Pan troglodytes), bonobo (Pan paniscus), gorilla (Gorilla gorilla), and orangutan (Pongo abelii), illustrates the key features of a phylogenetic tree such as nodes, branches, bootstrap numbers, and the scale.

Table 1.

List of Forensically Important Fly Species Commonly Collected in Korea

Family	Subfamily	Species	Genbank
Fanniidae		Fannia prisca	JX861413-7
Muscidae	Azelinae	Hydrotaea dentipes	JX861420-JX861429
		Hydrotaea occulta	JX861430
		Ophyra chalcogaster	JX861452-JX861456
		Ophyra leucostoma	JX861457-JX861459
		Ophyra nigra	JX861460-JX861468
		Muscina angustifrons	JX861436-JX861444
		Muscina stabulans	JX861449-JX861451
		Muscina pascuorum	JX861445-JX861448
	Muscinae	Musca domestica	JX861431-JX861435
	Phaoniinae	Phaonia aurea	JX861481&JX861482
Calliphoridae	Calliphorinae	Adrichina grahami	EU880180-EU880182
		Calliphora lata	EU880183-EU880187
		Calliphora vicina	EU880188-EU880192
		Triceratopyga calliphoroides	EU880176-EU880179
	Chrysomyinae^∗	Chrysomya megacephala	KF037969
		Chrysomya pinguis	FJ195381
		Phormia regina	JN257239&FJ360867
	Luciliinae	Hemipyrellia ligurriens	EU880206&EU880207
		Lucilia ampullaceae	EU925394
		Lucilia caesar	EU880193-EU880196
		Lucilia illustris	EU880197-EU880205
		Phaenicia sericata	EU880208-EU880212
Sarcophagidae	Sarcophaginae	Sarcophaga haemorrhoidalis	JX861406-JX861408
		Boettcherisca peregrina	JX861409-JX861412
		Helicophagella melanura	JX861418&JX861419
		Parasarcophaga albiceps	JX861469-JX861473
		Parasarcophaga harpax	JX861474&JX861475
		Parasarcophaga similis	JX861476-JX861480

^∗ Because the COI sequences of Korean Chrysomyinae flies were not submitted to NCBI Genbank yet, the listed accession numbers are those of conspecifics of which sequences are consistent with Korean counterparts.

Table 2.

Universal Primer Sequences for COI^∗

Name	Sequence	Binding site
F1 5	5′ -CCTTTAGAATTGCAGTCTAATGTCA-3′	tRNA-cysteine
F2 5	5′ -GGAGGATTTGGAAATTGATTAGTTCC-3′	′220-245 on COI
F3 5	5′ -CTGCTACTTTATGAGCTTTAGG-3′	1000-1022 on COI
R1 5	5′ -CCTAAATTTGCTCATGTTGACA-3′	2-23 on COII^†
R2 5	5′ -CAAGTTGTGTAAGCATC-3′	1327-1343 on COI
R3 5	5′ -CCAAAGAATCAAAATAAATGTTG-3′	688-710 on COI

^∗ COI: Cytochrome c oxidase subunit I;

^† COII: Cytochrome c oxidase subunit II

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