Soft Fruit Traceability in Food Matrices using Real-Time PCR

Palmieri, L.; Bozza, E.; Giongo, L.

doi:10.3390/nu1020316

Cited by 46 publications

(31 citation statements)

References 29 publications

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“…The use of the real‐time PCR method in food analysis in recent years has concentrated especially the on quantitative detection of the origins of vegetal and animal tissues in complex food mixtures (Palmieri and others 2009; Köppel and others 2011). In this context, the TaqMan probe method in the real‐time PCR technique is a very powerful technique in terms of both specifity and sensitivity in species detection.…”

Section: Discussionmentioning

confidence: 99%

Detection of Chicken and Turkey Meat in Meat Mixtures by Using Real‐Time PCR Assays

Kesmen

Yetiman

Şahi̇n

et al. 2012

Journal of Food Science

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In this study, TaqMan-based real-time Polymerase Chain Reaction (PCR) techniques were developed for the detection of chicken and turkey meat in raw and heat-treated meat mixtures. Primers and TaqMan probe sets were designed to amplify 86 bp and 136 bp fragments for the chicken and turkey species, respectively, on the mitochondrial NADH dehydrogenase subunit 2 gene. In the results, it was possible to detect each species at the level of 0.1 pg template DNA with the TaqMan probe technique without any cross-reactivity with nontarget species (bovine, ovine, donkey, pork, and horse) while the detection level was 1 pg template DNA using conventional PCR. The TaqMan probe assays used in this study allowed the detection of as little as 0.001% level of both species in the experimental meat mixtures, prepared by mixing chicken and turkey meat with beef at different levels (0.001% to 10%). In conclusion, TaqMan probe assays developed in this research are promising tools in the specific identification and sensitive quantification of meat species even in the case of heat-treated meat products, and suitable for a rapid, automated, and routine analysis.

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Section: Discussionmentioning

confidence: 99%

Detection of Chicken and Turkey Meat in Meat Mixtures by Using Real‐Time PCR Assays

Kesmen

Yetiman

Şahi̇n

et al. 2012

Journal of Food Science

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“…Primers for the first round of PCR were standard DNA barcoding primers with Illumina overhang sequences appended (Kozich et al, 2013). For rbcL we used the primers rbcL2, which binds near the 5′ end of the rbcL gene (Palmieri et al, 2009), and rbcLa-R, which binds near the middle of the rbcL gene (Kress and Erickson, 2007), which yield a PCR product of ~500 bp. For ITS2 we used the primers ITS2 S2F and ITS2 S3R, which bind to 5.8S and near the 3′ end of ITS2, respectively, and yield a PCR product of 350-400 bp (Chen et al, 2010).…”

Section: Methodsmentioning

confidence: 99%

How Many Plant Species are There, Where are They, and at What Rate are They Going Extinct?

Pimm

Joppa

2015

Annals of the Missouri Botanical Garden

178

104

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“…Primers for the first round of PCR were standard DNA barcoding primers with Illumina overhang sequences appended. For rbcL, we used the primers rbcL2 (Palmieri, Bozza, & Giongo, 2009) and rbcLa-R (Kress & Erickson 2007). For ITS2, we used the primers ITS2 S2F and ITS2 S3R (Chen et al, 2010).…”

Section: Dna Isolation and Sequencingmentioning

confidence: 99%

Quantitative and qualitative assessment of pollen DNA metabarcoding using constructed species mixtures

et al. 2018

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Pollen DNA metabarcoding-marker-based genetic identification of potentially mixed-species pollen samples-has applications across a variety of fields. While basic species-level pollen identification using standard DNA barcode markers is established, the extent to which metabarcoding (a) correctly assigns species identities to mixes (qualitative matching) and (b) generates sequence reads proportionally to their relative abundance in a sample (quantitative matching) is unclear, as these have not been assessed relative to known standards. We tested the quantitative and qualitative robustness of metabarcoding in constructed pollen mixtures varying in species richness (1-9 species), taxonomic relatedness (within genera to across class) and rarity (5%-100% of grains), using Illumina MiSeq with the markers rbcL and ITS2. Qualitatively, species composition determinations were largely correct, but false positives and negatives occurred. False negatives were typically driven by lack of a barcode gap or rarity in a sample. Species richness and taxonomic relatedness, however, did not strongly impact correct determinations. False positives were likely driven by contamination, chimeric sequences and/or misidentification by the bioinformatics pipeline. Quantitatively, the proportion of reads for each species was only weakly correlated with its relative abundance, in contrast to suggestions from some other studies. Quantitative mismatches are not correctable by consistent scaling factors, but instead are context-dependent on the other species present in a sample. Together, our results show that metabarcoding is largely robust for determining pollen presence/absence but that sequence reads should not be used to infer relative abundance of pollen grains.

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Soft Fruit Traceability in Food Matrices using Real-Time PCR

Cited by 46 publications

References 29 publications

Detection of Chicken and Turkey Meat in Meat Mixtures by Using Real‐Time PCR Assays

Detection of Chicken and Turkey Meat in Meat Mixtures by Using Real‐Time PCR Assays

How Many Plant Species are There, Where are They, and at What Rate are They Going Extinct?

Quantitative and qualitative assessment of pollen DNA metabarcoding using constructed species mixtures

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