title: A multiplex real-time PCR method for the quantification of beef and pork fractions in minced meat.
abstract: One popular staple food in many lands is minced meat, traditionally prepared from beef and/or pork fractions. While beef is the more expensive of the two meat fractions, the possibility exists for manufacturers to fraudulently declare higher proportions of it. Additionally, the need exists to protect consumers who, out of medical or ethical reasons, reject specific meat fractions. In this work, we report on a quantitative triplex real-time PCR approach for the quantification of meat in minced meat products. With the method, beef and pork fractions are quantified employing primer and probe sequences that specifically recognise cow and pig components, against the backdrop of myostatin, a universal sequence commonly found in mammals and poultry species. The limit of detection of the qPCR method was 20 genome equivalents, while the measurement of uncertainty was determined at 1.83%. The method was validated on several commercially available minced meat products and performed well in terms of handling, reproducibility and robustness.
authors.affiliation: Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Germany. Electronic address: azuka.iwobi@lgl.bayern.de.
authors.forename: A
authors.initials: A
authors.lastname: Iwobi
authors.affiliation: Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Sebah
authors.affiliation: Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Germany.
authors.forename: I
authors.initials: I
authors.lastname: Kraemer
authors.affiliation: Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Losher
authors.affiliation: Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Germany.
authors.forename: G
authors.initials: G
authors.lastname: Fischer
authors.affiliation: Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Germany.
authors.forename: U
authors.initials: U
authors.lastname: Busch
authors.affiliation: Bavarian Health and Food Safety Authority, Veterinaerstrasse 2, 85764, Germany.
authors.forename: I
authors.initials: I
authors.lastname: Huber
mesh_terms: D000818:Animals; D002417:Cattle; D005260:Female; D057230:Limit of Detection; D008460:Meat / Q000032:analysis*; D008461:Meat Products / Q000032:analysis*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D013552:Swine
keywords: Beef; Pork; Real-time PCR; Species identification and quantification; Validation