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Development of a combined selective enrichment method and polymerase chain reaction (PCR) assay for sensitive detection of Salmonella in food samples
Authors:Marsiglia  M.L.  Ikuta  N.  Fonseca  A.K.  Schuch  D.T.  Ho¨tzel  I.  Ozaki  L.S.  Marques  E.K.  Lunge  V.R.
Affiliation:(1) Departamento de Biotecnologia, Instituto de Biocie^ncias – UFRGS, Av. Bento Gonc¸alves, 9500, Caixa Postal, 15005, 91501-970 Porto Alegre, RS, Brazil;(2) Simbios Biotecnologia, Av. Bento Gonc¸alves, 9500, Pre´dio 43421, Caixa Postal 15092, 91501-970 Porto Alegre, RS, Brazil;(3) Laborato´rio de Refere^ncia Animal (LARA-RS), Estrada da Ponta Grossa 3036, Porto Alegre, RS, Brazil
Abstract:PCR assays were formatted using primer pairs homologous to phoE and invA genes. The amplification conditions were optimized with pure cultures and reactions were carried out to define selectivity, specificity and sensitivity of both primer pairs. The performance of the invA primer pair was better than that of the phoE pair, making the specific detection of Salmonella serovars and strains isolated from different food samples possible. Using the invA primer pair, the combined selective enrichment method with the polymerase chain reaction assay was established and used to detect Salmonella from artificially multi-contaminated food samples. The complete procedure detected as few as three cells of Salmonella (3 c.f.u.) from milk and meat samples.
Keywords:Food microbiology  polymerase chain reaction  Salmonella detection
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