Universal primers for species authentication of animal foodstuff in a single polymerase chain reaction

Abstract

These primers permit the identification of most animal taxa employed in human nutrition, from invertebrates such as molluscs to higher vertebrates, distinguishing between species of the same genus. The short fragment amplified within the 16S rDNA exhibits phylogenetic value and could be considered universal based on the wide taxonomic range assayed. The primers are easy to use and accessible for laboratories with a modest budget, as well as being valuable for consumer information and to reveal food fraud.

Authentication of commercial candy ingredients using DNA PCR‐cloning methodology

“…5 and Table 2). More animal species were found from DNA than from labels with Horreo et al 18 primers, whereas for plants it was the opposite, with more plants and fruits stated in the labels than found from DNA with Han et al 19 primers.…”

“…Since candies are highly processed their DNA is likely degraded, 17 and therefore we targeted short species-specific sequences. For animal species we employed the primers 16S-HF 5 ′ -ATAACACGAGAAGACCCT-3 ′ and 16S-HR 5 ′ -CCCRCGGTCGCCCCAAC-3 ′ developed by Horreo et al 18 that amplify an 80-122 base pair (bp) fragment within the 16S rRNA gene. PCR reaction was performed with: 5 μL DNA extraction from the candy (from less than 0.0005 ng μL −1 to 0.328 ng μL −1 ), 0.125 μL Taq polymerase from the PCR Core Kit Plus, polymerase solution with Mg 2+ 1×, 0.5 μL of each primer (10 μmol L −1 ), 0.5 μL dideoxynucleotides (dNTPs with U), 0.5 μL Uracil glycosylase and bidistilled water up to 25 μL of total volume.…”

‘Representative Genes’, is it OK to use a small amount of data to obtain a phylogeny that is at least close to the true tree?

“…Nonphylogenetic signal may become dominant and yield incongruent, yet statistically highly supported phylogenomic trees (Jeffroy et al, 2006), and hence it could be useful to know the existence of one gene in each group of species (such as family or genus) that would be representative of its evolution, being especially useful to construct the phylogenetic relationships of this group. In this sense, Horreo et al (2012) have found recently a very short fragment of the 16S rDNA (around 120 bp) with some phylogenetic value.…”

“…In this sense, Horreo et al . () have found recently a very short fragment of the 16S rDNA (around 120 bp) with some phylogenetic value.…”

Advances in Authenticity Testing for Meat Speciation

“…Horreo et al (2013) designed universal primers based on the nucleotide sequences of the 16S rRNA gene of 40 animal species that belonged to seven different taxonomic classes (Actinopterygii, Gasteropoda, Cephalaspidomorphi, Amphibia, Chondrichthyes, Aves, and Mammalia), obtaining PCR amplicons ranging from 80 to 125 bp. Sequencing analyses confirmed the differences among species and, although very short, they were considered very informative and enabled clustering of samples into their respective taxonomic group.…”