Desenvolvimento de um método seguro pela técnica de PCR em tempo real para detecção de dez espécies animais em produtos cárneos

• Ano de defesa: 2017
• Autor(a) principal: Lissandra Sousa Dalsecco
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Minas Gerais; UFMG
• Palavras-chave em Português: Substituição de espécies; Rotulagem inadequada; Produtos cárneos; PCR em Tempo Real; Alimentos de origem animal Análise; Alimentos de origem animal Adulteração e inspeção; Rotulagem Inspeção; Carne Inspeção; Reação em cadeia de polimerase
• Link de acesso: http://hdl.handle.net/1843/BUOS-BBYQTX

Species identification in food is a subject of significant interest for the entire production chain, for government inspection and in academic circles. Fraud by addition or substitution of species is against the law in Brazil and can lead to serious consequences, including economic losses and public health risks. Several methods are available for identifying the origin of food and, among these, the DNA techniques have proven to be practical, reliable, specific and highly sensitive for fraud detection. The present work is aimed at the development of a test to detect DNA from Bos taurus, Sus scrofa, Ovis aries, Capra hircus, Gallus gallus, Meleagris gallopavo, Bubalus bubalis, Equus caballus, Felis catus and Canis familiaris in meat products. The method is based on real-time PCR and combines the use of universal primers (for internal positive control) and species-specific primers for the detection of target DNA. After each reaction, the PCR products are subject to melt curve analysis to check if the amplified DNA corresponds to the expected fragment of each species. Method precision was evaluated on 45 experimental meat mixtures in proportions of 50-50% or 99-1% plus one mixture with 10% of each species. The test correctly detected the species in each mixture, including those present in 1% of the total content. The method was then applied to the analysis of 14 commercial meat products and the results revealed that six of these had non-declared bovine and/or chicken material. The experimental mixtures and commercial products analyses were carried out in the two participating laboratories (LGEV and Myleus), obtaining 100% reproducibility. The developed test proved to be effective and reliable for a routine of species detection in meat products