Development of a real-time PCR protocol for the species origin confirmation of isolated animal particles detected by NIRM.
At present, European legislation prohibits totally the use of processed animal proteins in feed for all farmed animals (Commission Regulation (EC) No. 1234/2003-extended feed ban). A softening of the feed ban for non-ruminants would nevertheless be considered if alternative methods could be used to gain more information concerning the species origin of processed animal proteins than that which can be provided by classical optical microscopy. This would allow control provisions such as the ban of feeding animals with proteins from the same species or intra-species recycling (Regulation (EC) No. 1774/2002). Two promising alternative methods, near-infrared microscopy (NIRM) and real-time polymerase chain reaction (PCR), were combined to authenticate, at the species level, the presence of animal particles. The paper describes the improvements of the real-time PCR method made to the DNA extraction protocol, allowing five PCR analyses to be performed with the DNA extracted from a single particle.
Département Valorisation des Productions Agricoles, Centre Wallon de Recherches Agronomiques, Bâtiment Henseval, Chaussée de Namur, 24, B-5030 Gembloux, Belgium. fumiere@cra.wallonie.be
O
O
Fumière
A
A
Marien
J A
JA
Fernández Pierna
V
V
Baeten
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G
Berben
D053000:Analytic Sample Preparation Methods; D000821:Animal Feed / Q000032:analysis* / Q000592:standards; D000822:Animal Husbandry / Q000331:legislation & jurisprudence / Q000379:methods; D000818:Animals; D000829:Animals, Domestic / Q000235:genetics; D004247:DNA / Q000302:isolation & purification; D004044:Dietary Proteins / Q000592:standards; D005062:European Union; D005506:Food Contamination / Q000517:prevention & control; D005513:Food Inspection / Q000191:economics / Q000379:methods*; D005517:Foodborne Diseases / Q000517:prevention & control; D005838:Genotype; D007220:Industrial Waste / Q000032:analysis / Q000191:economics; D008461:Meat Products / Q000032:analysis*; D008462:Meat-Packing Industry / Q000191:economics / Q000379:methods; D008853:Microscopy / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D013045:Species Specificity; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*
D004044:Dietary Proteins; D007220:Industrial Waste; D004247:DNA