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Molecular identification of Salmonella enterica subsp. enterica serovar Gallinarum biovars Gallinarum and Pullorum by a duplex PCR assay

Batista, Diego Felipe Alves, de Freitas Neto, Oliveiro Caetano, de Almeida, Adriana Maria, Barrow, Paul Andrew, de Oliveira Barbosa, Fernanda, Berchieri Junior, Angelo
DNA, Salmonella enterica subsp. enterica, biosecurity, pathogens, polymerase chain reaction, poultry, serotypes
Salmonella enterica subsp. enterica serovar Gallinarum biovar Gallinarum (S. Gallinarum) and biovar Pullorum (S. Pullorum) are 2 poultry pathogens that cause major economic losses to the poultry industry worldwide. Control of both diseases mainly relies on the adoption of biosecurity programs, and success is dependent on accurate and fast detection. Based on this concept, we developed a duplex PCR assay, targeting 2 chromosomal sequences, which allowed us to precisely identify and differentiate S. Gallinarum and S. Pullorum field strains. This assay was validated by testing genomic DNA from 40 S. Gallinarum and 29 S. Pullorum field strains, 87 other Salmonella serovars, and 7 non-Salmonella strains. The serovar identifier region (SIR) primers produced a fragment only in S. Gallinarum and S. Pullorum strains, whereas the fragment from the ratA coding sequence, which was previously demonstrated to differentiate the 2 biovars, was also amplified from other Salmonella serovars. Our results showed that the combination of both SIR and ratA amplifications could be used to identify as well as to differentiate colonies of S. Gallinarum and S. Pullorum reliably. Thus, we believe this methodology can be a useful ancillary tool for routine veterinary diagnostic laboratories by providing rapid, accurate results.