Applicability assessment of a real-time PCR assay for the specific detection of bovine,ovine and caprine material in feedstuffs |
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Authors: | Nicolette Pegels Isabel González Irene Martín María Rojas Teresa García Rosario Martín |
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Affiliation: | 1. The State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, No. 22 Hankou Rd, Gulou District, Nanjing 210093, PR China;2. School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, PR China;1. Department of Tourism and Environmental Sciences, Shaanxi Normal University, Xi''an 710062 (China);2. Key Laboratory of Disaster Survey and Mechanism Simulation of Shaanxi Province, Baoji University of Arts and Sciences, Baoji 721013 (China);3. State Key Laboratory of Soil Erosion and Dryland Farming on the Loess Plateau, Institute of Soil and Water Conservation, Chinese Academy of Sciences, Yangling 712100 (China);1. CINSO – (CITEDEF), UNIDEF (Strategic I & D for Defense)-CONICET-Ministry of Defense, Juan Bautista de la Salle 4970, B1603ALO Villa Martelli, Buenos Aires, Argentina;2. The National Council for Scientific and Technical Research (CONICET), Rivadavia 1917, C1033AAJ, Buenos Aires, Argentina;3. Food Technology Institute-INTA, Morón, B1708WAB, Buenos Aires, Argentina;4. National University of Luján, Route 5 y Avenida Constitución – Luján 6700, Buenos Aires, Argentina;1. Groupe de Recherche en Génie Rural, Earth and Life Institute—Environmental Sciences, Université catholique de Louvain, Croix du Sud 2 (L7.05.02), Louvain-la-Neuve 1348, Belgium;2. Groupe de Recherche en Physiologie Végétale, Earth and Life Institute—Agronomy, Université catholique de Louvain, Croix du Sud 4-5 (L7.07.13), Louvain-la-Neuve 1348, Belgium;3. Groupe de Recherche en Sciences du Sol, Earth and Life Institute—Environmental Sciences, Université catholique de Louvain, Croix du Sud 2 (L7.05.10), Louvain-la-Neuve 1348, Belgium;1. Department of Mathematics, Bohai University, Jinzhou 121013, China;2. Research Institute of Food Science, Bohai University, Jinzhou 121013, China;3. College of Science, Zhejiang Sci-Tech University, Hangzhou 310018, China;4. Department of Applied Mathematics, Shanghai Institute of Technology, Shanghai 201418, China;5. College of Chemistry, Chemical Engineering and Food Safety, Bohai University, Jinzhou 121013, China |
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Abstract: | A TaqMan real-time polymerase chain reaction (PCR) method was developed for specific detection of bovine, ovine and caprine processed animal protein (PAP) in industrial feedstuffs. The method uses species-specific primers and probes targeting short mitochondrial D-loop sequences, and a positive amplification control based on 18S rRNA gene. The applicability of the real-time PCR protocol was assessed through analysis of 126 industrial feed samples that were manufactured to reproduce rendering processes of commercial feeds destined for farmed animals. The assay successfully classified samples as positive or negative according to the ruminant composition, enabling qualitative detection of banned material in feeds at levels as low as 0.1%. Although the method provides quantitative potential, results suggest that the real quantitative capability of the assay is limited by the existing variability in terms of composition and processing treatments of the feeds, which affect the amount and quality of amplifiable DNA. |
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