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Development of a Rapid Real-Time PCR Method as a Tool To Quantify Viable Photobacterium phosphoreum Bacteria in Salmon (Salmo salar) Steaks ArchiMer
Mace, Sabrina; Mamlouk, Kelthoum; Chipchakova, Stoyka; Prevost, Herve; Joffraud, Jean-jacques; Dalgaard, Paw; Pilet, Marie-france; Dousset, Xavier.
A specific real-time PCR quantification method combined with a propidium monoazide sample treatment step was developed to determine quantitatively the viable population of the Photobacterium phosphoreum species group in raw modified-atmosphere-packed salmon. Primers were designed to amplify a 350-bp fragment of the gyrase subunit B gene (gyrB) of P. phosphoreum. The specificity of the two primers was demonstrated by using purified DNA from 81 strains of 52 different bacterial species. When these primers were used for real-time PCR in pure culture, a good correlation (R-2 of 0.99) was obtained between this method and conventional enumeration on marine agar (MA). Quantification was linear over 5 log units as confirmed by using inoculated salmon samples. On...
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Ano: 2013 URL: https://archimer.ifremer.fr/doc/00135/24623/22772.pdf
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