In this work four species-specific primers and probes were designed and evaluated for the detection and quantification of bovine, ovine, swine and chicken mitochondrial DNA in feeds. PCR primers were optimized using conventional and Real Time PCR, to detect short species-specific sequences amplifiable from heat treated material. Both methods confirmed the high specificity of the primers designed. Real time quantitative PCR assay allowed the detection of as few as 0.01 ng and 0.05 ng of ovine and bovine genomic DNA, respectively. The detection limit for swine and chicken genomic DNA was 0.5 ng. Sensitivity levels observed in DNA extracted from meat samples processed according to EU legislation were different compared to those in genomic DNAs previously described. They resulted in swine 5 fg of MBM DNA, in chicken 25 ng, in ovine and bovine 50 ng. We confirmed the efficiency and specificity of primers in RT-PCR to detect 0.5% of bovine, ovine, swine and chicken MBM in contaminated feedstuffs. (C) 2007 Elsevier Ltd. All rights reserved.
Frezza, D., Giambra, V., Chegdani, F., Fontana, C., Maccabiani, G., Losio, N., et al. (2008). Standard and Light-Cycler PCR methods for animal DNA species detection in animal feedstuffs. INNOVATIVE FOOD SCIENCE & EMERGING TECHNOLOGIES, 9(1), 18-23 [10.1016/j.ifset.2007.04.008].
Standard and Light-Cycler PCR methods for animal DNA species detection in animal feedstuffs
FREZZA, DOMENICO;
2008-01-01
Abstract
In this work four species-specific primers and probes were designed and evaluated for the detection and quantification of bovine, ovine, swine and chicken mitochondrial DNA in feeds. PCR primers were optimized using conventional and Real Time PCR, to detect short species-specific sequences amplifiable from heat treated material. Both methods confirmed the high specificity of the primers designed. Real time quantitative PCR assay allowed the detection of as few as 0.01 ng and 0.05 ng of ovine and bovine genomic DNA, respectively. The detection limit for swine and chicken genomic DNA was 0.5 ng. Sensitivity levels observed in DNA extracted from meat samples processed according to EU legislation were different compared to those in genomic DNAs previously described. They resulted in swine 5 fg of MBM DNA, in chicken 25 ng, in ovine and bovine 50 ng. We confirmed the efficiency and specificity of primers in RT-PCR to detect 0.5% of bovine, ovine, swine and chicken MBM in contaminated feedstuffs. (C) 2007 Elsevier Ltd. All rights reserved.File | Dimensione | Formato | |
---|---|---|---|
2008 DF Feed staff LC IFSET.pdf
accesso aperto
Descrizione: Articolo scientifico sperimentale
Dimensione
398.41 kB
Formato
Adobe PDF
|
398.41 kB | Adobe PDF | Visualizza/Apri |
I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.