Development of a real time PCR protocol for the species origin confirmation of isolated animal particles detected by NIRM
International audience ; At present, the European legislation prohibits totally the use of processed animal proteins (PAPs) in feed for all farmed animals [Commission Regulation (EC) n°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 on the species origin of PAPs than what can be provided by classical optical microscopy. This would allow to control provisions such as the ban of feeding animals with proteins from the same species or intra-species recycling [Regulation (EC) n°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 present paper describes the improvements of the real-time PCR method lying in the DNA extraction protocol and allowing to perform up to 5 PCR analyses with the DNA extracted from a single particle.
