A multiplex assay to identify 18 european mammal species from mixtures using the mitochondrial cytochrome b gene

Shanan S. Tobe, Adrian M.T. Linacre

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

A novel species-specific multiplex to identify 18 common European mammalian species (badger, cat, cow, dog, donkey, fox, goat, guinea pig, harvest mouse, hedgehog, horse, house mouse, human, pig, rabbit, rat, red deer and sheep), many of which are often associated with forensic investigations, has been developed. The assay is based on the mitochondrial cytochrome b gene, which is commonly used in species identification and phylogeny studies. Areas of homology and variation were identified and were used to create universal and species-specific primers. The species-specific primers were designed such that they will only react with the species for which they were designed. Two primer sets were designed for each species making the test self-confirmatory. All primer sets produced the expected results. The multiplex was balanced at template concentration of 40 000 copies (approximately 1.36 pg). Validation was accomplished by analysing the same sample ten times to determine run variation and several samples for each species to determine between-sample variation. Twenty-eight additional mammalian species were reacted with the multiplex. The multiplex provides, for the first time, a definitive method for identification of species in a forensic context.
LanguageEnglish
Pages340-347
Number of pages8
JournalElectrophoresis
Volume29
Issue number2
DOIs
Publication statusPublished - Jan 2008

Fingerprint

Cytochromes b
Mammals
Rats
Assays
Genes
Mustelidae
Deer
Hedgehogs
Equidae
Phylogeny
Goats
Horses
Sheep
Guinea Pigs
Cats
Swine
Dogs
Rabbits

Keywords

  • cytochrome b gene
  • forensic science
  • mixtures
  • species identification
  • trace samples

Cite this

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title = "A multiplex assay to identify 18 european mammal species from mixtures using the mitochondrial cytochrome b gene",
abstract = "A novel species-specific multiplex to identify 18 common European mammalian species (badger, cat, cow, dog, donkey, fox, goat, guinea pig, harvest mouse, hedgehog, horse, house mouse, human, pig, rabbit, rat, red deer and sheep), many of which are often associated with forensic investigations, has been developed. The assay is based on the mitochondrial cytochrome b gene, which is commonly used in species identification and phylogeny studies. Areas of homology and variation were identified and were used to create universal and species-specific primers. The species-specific primers were designed such that they will only react with the species for which they were designed. Two primer sets were designed for each species making the test self-confirmatory. All primer sets produced the expected results. The multiplex was balanced at template concentration of 40 000 copies (approximately 1.36 pg). Validation was accomplished by analysing the same sample ten times to determine run variation and several samples for each species to determine between-sample variation. Twenty-eight additional mammalian species were reacted with the multiplex. The multiplex provides, for the first time, a definitive method for identification of species in a forensic context.",
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A multiplex assay to identify 18 european mammal species from mixtures using the mitochondrial cytochrome b gene. / Tobe, Shanan S.; Linacre, Adrian M.T.

In: Electrophoresis, Vol. 29, No. 2, 01.2008, p. 340-347.

Research output: Contribution to journalArticle

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