1. Molecular detection and quantification of the protozoan Bonamia ostreae in the flat oyster, Ostrea edulis
- Author
-
Sylvie Ferrand, Inmaculada López-Flores, Maeva Robert, Isabelle Arzul, Bruno Chollet, Celine Garcia, Jean-Pierre Joly, and Cyrille Francois
- Subjects
Oyster ,Ostrea edulis ,Haplosporida ,Melting temperature ,Protozoan Proteins ,Bonamia ostreae ,Polymerase Chain Reaction ,Host-Parasite Interactions ,law.invention ,03 medical and health sciences ,law ,Quantification ,biology.animal ,Ostrea ,Heart imprints ,Animals ,Molecular Biology ,Polymerase chain reaction ,030304 developmental biology ,0303 health sciences ,biology ,Ecology ,Reproducibility of Results ,04 agricultural and veterinary sciences ,Cell Biology ,DNA, Protozoan ,biology.organism_classification ,Molecular biology ,Peptide Fragments ,Detection ,Real time PCR ,Real-time polymerase chain reaction ,Connexin 43 ,Specific primers ,Linear Models ,040102 fisheries ,0401 agriculture, forestry, and fisheries ,Polymorphism, Restriction Fragment Length - Abstract
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
- Published
- 2009
- Full Text
- View/download PDF