Top

Published in:

01-09-2007 | Short Communication

Two universal primer sets for species identification among vertebrates

Authors: Takashi Kitano, Kazuo Umetsu, Wei Tian, Motoki Osawa

Published in: International Journal of Legal Medicine | Issue 5/2007

Abstract

The aim of this study was to develop a simple method using universal primers for species identification based on direct PCR sequencing. Two primer sets were designed based on the conserved regions of the 12S and 16S rRNA loci detected by the comprehensive sequence comparison among 30 mammalian whole mitochondrial genomes. In humans, the expected sizes of PCR products of the 12S and 16S rRNAs were 215 and 244 bp, respectively. Both primer sets successfully amplified the expected PCR products from various kinds of vertebrates including mammals, birds, reptiles, amphibians, and fish, and the sequenced segments contained sufficient nucleotide differences to identify each animal species. A case example of the identification of a piece of buried bone of unknown species is presented, and the species was identified as a pig by this method.

Available only for authorised users

Zehner R, Zimmermann S, Mebs D (1998) RFLP and sequence analysis of the cytochrome b gene of selected animals and man: methodology and forensic application. Int J Legal Med 111:323–327PubMedCrossRef

Bravi CM, Liron JP, Mirol PM, Ripoli MV, Peral-Garcia P, Giovambattista G (2004) A simple method for domestic animal identification in Argentina using PCR-RFLP analysis of cytochrome b gene. Leg Med (Tokyo) 6:246–251

Kocher TD, Thomas WK, Meyer A, Edwards SV, Pääbo S, Villablanca FX, Wilson AC (1989) Dynamics of mitochondrial DNA evolution in animals: amplification and sequencing with conserved primers. Proc Natl Acad Sci USA 86:6196–6200PubMedCrossRef

Parson W, Pegoraro K, Niederstatter H, Föger M, Steinlechner M (2000) Species identification by means of the cytochrome b gene. Int J Legal Med 114:23–28PubMedCrossRef

Hsieh HM, Chiang HL, Tsai LC, Lai SY, Huang NE, Linacre A, Lee JC (2001) Cytochrome b gene for species identification of the conservation animals. Forensic Sci Int 122:7–18CrossRef

Branicki W, Kupiec T, Pawlowski R (2003) Validation of cytochrome b sequence analysis as a method of species identification. J Forensic Sci 48:83–87PubMed

Bellis C, Ashton KJ, Freney L, Blair B, Griffiths LR (2003) A molecular genetic approach for forensic animal species identification. Forensic Sci Int 134:99–108PubMedCrossRef

Balitzki-Korte B, Anslinger K, Bartsch C, Rolf B (2005) Species identification by means of pyrosequencing the mitochondrial 12S rRNA gene. Int J Legal Med 119:291–294PubMedCrossRef

Bataille M, Crainic K, Leterreux M, Durigon M, de Mazancourt P (1999) Multiplex amplification of mitochondrial DNA for human and species identification in forensic evaluation. Forensic Sci Int 99:165–170PubMedCrossRef

10.

Dalmasso A, Fontanella E, Piatti P, Civera T, Rosati S, Bottero MT (2004) A multiplex PCR assay for the identification of animal species in feedstuffs. Mol Cell Probes 18:81–87PubMedCrossRef

11.

Thompson JD, Gibson TJ, Higgins DG (1994) ClustalW: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680PubMedCrossRef

12.

Nei M (1987) Molecular evolutionary genetics. Columbia University Press, New York, pp 254–286

13.

Kimura M (1980) A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16:111–120PubMedCrossRef

14.

Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425PubMed

15.

Springer MS, Douzery E (1996) Secondary structure and patterns of evolution among mammalian mitochondrial 12S rRNA molecules. J Mol Evol 43:357–373PubMed

16.

Burk A, Douzery EJP, Springer MS (2002) The secondary structure of mammalian mitochondrial 16S rRNA molecules: refinements based on a comparative phylogenetic approach. J Mamm Evol 9:225–252CrossRef

Title: Two universal primer sets for species identification among vertebrates
Authors: Takashi Kitano
Kazuo Umetsu
Wei Tian
Motoki Osawa
Publication date: 01-09-2007
Publisher: Springer-Verlag
Published in: International Journal of Legal Medicine / Issue 5/2007
Print ISSN: 0937-9827
Electronic ISSN: 1437-1596
DOI: https://doi.org/10.1007/s00414-006-0113-y

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Two universal primer sets for species identification among vertebrates

Abstract

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 5/2007

Increase in clusterin-containing follicles in the adenohypophysis of drug abusers

Hungarian mtDNA population databases from Budapest and the Baranya county Roma

Soft tissue removal by maceration and feeding of Dermestes sp.: impact on morphological and biomolecular analyses of dental tissues in forensic medicine

Carisoprodol intoxications: a retrospective study of forensic autopsy material from 1992–2003

Neuronal and glial apoptosis in human traumatic brain injury

Y-chromosomal microsatellite mutation rates in a population sample from northwestern Germany