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Molecular detection and quantification of the protozoan Bonamia ostreae in the flat oyster, Ostrea edulis
Provedor de dados: 5
Autores: Robert, Maeva; Garcia, Celine; Chollet, Bruno; Lopez-flores, Inmaculada; Ferrand, Sylvie; Francois, Cyrille; Joly, Jean-pierre; Arzul, Isabelle.
Bonamia ostreae is an intracellular protozoan which is recognized as a cause of mortality in European populations of flat oysters (Ostrea edulis). Based on the recent characterization of actin genes of B. ostreae, specific primers were designed for real-time PCR using SYBR® Green chemistry. Specificity was demonstrated by the unique melting temperature peak observed in positive samples and by the lack of amplification in samples of oysters infected by closely related parasites, including Bonamia exitiosa. A calibration curve using a cloned template was defined to estimate copy number. The assay had a 6 log- dynamic range, mean inter- and intra-assay variation coefficients of &lt;1% and a minimum detection limit of 50 gene copies per reaction. Using...
Tipo: Text
Palavras-chave: Heart imprints; Ostrea edulis; Quantification; Detection; Real time PCR; Bonamia ostreae.
Ano: 2009
URL: http://archimer.ifremer.fr/doc/2009/publication-6934.pdf