Polymerase chain reaction-based analysis to detect terrestrial animal protein in fish meal.
The recent European bovine spongiform encephalopathy crisis has focused attention on the importance of adopting stringent control measures to avoid the risk of the diffusion of mad cow disease through meat meal-based animal feedstuffs. Potential adulteration of such feedstuffs with bone particles from terrestrial animals is determined by microscopic examination by law before the release of these feedstuffs for free circulation in the European Community. This study describes a DNA monitoring method to examine fish meal for contamination with mammalian and poultry products. A polymerase chain reaction (PCR) method based on the nucleotide sequence variation in the 12S ribosomal RNA gene of mitochondrial DNA was developed and evaluated. Three species-specific primer pairs were designed for the identification of ruminant, pig, and poultry DNA. The specificity of the primers used in the PCR was tested by comparison with DNA samples for several vertebrate species and confirmed. The PCR specifically detected mammalian and poultry adulteration in fish meals containing 0.125% beef, 0.125% sheep, 0.125% pig, 0.125% chicken, and 0.5% goat. A multiplex PCR assay for ruminant and pig adulteration was optimized and had a detection limit of 0.25%.
Bellagamba
Federica
F
Dipartimento di Scienze e Tecnologie Veterinarie per la Sicurezza Alimentare, Facoltà di Medicina Veterinaria, Università degli Studi di Milano, Via Trentacoste, 2, 20134 Milano, Italy. federica.bellagamba@unimi.it
Valfrè
Franco
F
Panseri
Sara
S
Moretti
Vittorio M
VM
D000821:Animal Feed / Q000032:analysis; D000818:Animals; D001483:Base Sequence; D001688:Biological Products; D002417:Cattle; D002645:Chickens; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000032:analysis*; D005396:Fish Products / Q000032:analysis*; D005506:Food Contamination / Q000032:analysis; D005784:Gene Amplification; D006041:Goats; D008903:Minerals; D016133:Polymerase Chain Reaction / Q000379:methods*; D011506:Proteins / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D012756:Sheep; D013045:Species Specificity; D013552:Swine
D001688:Biological Products; D017931:DNA Primers; D004272:DNA, Mitochondrial; D008903:Minerals; D011506:Proteins; C027762:bone meal