title: A multiplex PCR for the simultaneous detection of Tenacibaculum maritimum and Edwardsiella tarda in aquaculture.
abstract: A specific and sensitive multiplex PCR (mPCR) method was developed as a useful tool for the simultaneous detection of two important flatfish pathogens in marine aquaculture, Tenacibaculum maritimum and Edwardsiella tarda. In fish tissues, the average detection limit for these mPCR-amplified organisms was 2 × 10 ⁵ ± 0.2 CFU/g and 4 × 10 ⁵ ± 0.3 CFU/g, respectively. These values are similar or even lower than those previously obtained using the corresponding single PCR. Moreover, mPCR did not produce any nonspecific amplification products when tested against 36 taxonomically related and unrelated strains belonging to 33 different bacterial species. Large amounts of DNA from one of the target bacterial species in the presence of low amounts from the other did not have a significant effect on the amplification sensitivity of the latter.
authors.affiliation: Department of Microbiology and Parasitology, Faculty of Biology/CIBUS, University of Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: Nuria
authors.initials: N
authors.lastname: Castro
authors.affiliation: Department of Microbiology and Parasitology, Faculty of Biology/CIBUS, University of Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: Alicia E
authors.initials: AE
authors.lastname: Toranzo
authors.affiliation: Department of Microbiology and Parasitology, Faculty of Biology/CIBUS, University of Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Magariños
mesh_terms: D000818:Animals; D017756:Aquaculture; D020609:Edwardsiella tarda / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D004756:Enterobacteriaceae Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D005393:Fish Diseases / Q000175:diagnosis / Q000382:microbiology*; D005399:Fishes / Q000382:microbiology*; D045826:Flavobacteriaceae Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D060493:Tenacibaculum / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*
keywords: Edwardsiella tarda; Tenacibaculum maritimum; aquaculture; fish pathology; multiplex PCR