title: Development and validation of species-specific molecular diagnostic tool for Opisthorchis felineus (Digenea, Opisthorchiidae) metacercariae.
abstract: Opisthorchis felineus (family Opisthorchiidae) is a parasitic flatworm representing a serious threat to humans in some countries. Opisthorchiasis occurs after consumption of raw or undercooked cyprinid fish infected by the metacercarial stage of the parasite. Due to its small size, detection of the parasite in fish fillet is time-consuming and difficult. Furthermore, isolated metacercariae can be identified to genus but not to species level using morphological features and molecular techniques are necessary. In this work, we describe the development of primers for a diagnostic PCR amplification of a 254-bp fragment of the cytochrome c oxidase I in the mitochondrion of Opisthorchis felineus metacercariae isolated from fish fillet, together with a validation protocol for this method.
authors.affiliation: Department of Veterinary Medical Sciences, Alma Mater Studiorum University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, BO, Italy. Electronic address: monica.caffara@unibo.it.
authors.forename: Monica
authors.initials: M
authors.lastname: Caffara
authors.affiliation: Marine Microbiology Laboratory, Istituto Zooprofilattico Sperimentale del Piemonte Liguria e Valle d'Aosta, Via degli Stagnoni 96, 19136 La Spezia, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Serracca
authors.affiliation: Department of Veterinary Medical Sciences, Alma Mater Studiorum University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, BO, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Gustinelli
authors.affiliation: Marine Microbiology Laboratory, Istituto Zooprofilattico Sperimentale del Piemonte Liguria e Valle d'Aosta, Via degli Stagnoni 96, 19136 La Spezia, Italy.
authors.forename: Walter
authors.initials: W
authors.lastname: Vencia
authors.affiliation: Marine Microbiology Laboratory, Istituto Zooprofilattico Sperimentale del Piemonte Liguria e Valle d'Aosta, Via degli Stagnoni 96, 19136 La Spezia, Italy.
authors.forename: Irene
authors.initials: I
authors.lastname: Rossini
authors.affiliation: Fish Diseases Laboratory, Istituto Zooprofilattico Sperimentale del Piemonte Liguria e Valle d'Aosta, Via Bologna 148, Turin, Italy.
authors.forename: Marino
authors.initials: M
authors.lastname: Prearo
authors.affiliation: Department of Veterinary Medical Sciences, Alma Mater Studiorum University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia, BO, Italy.
authors.forename: Maria L
authors.initials: ML
authors.lastname: Fioravanti
mesh_terms: D000818:Animals; D003530:Cyprinidae / Q000469:parasitology; D017931:DNA Primers / Q000235:genetics; D005393:Fish Diseases / Q000175:diagnosis* / Q000469:parasitology; D058488:Metacercariae / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D009889:Opisthorchiasis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D009891:Opisthorchis / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D057089:Pathology, Molecular; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity
chemical_list: D017931:DNA Primers
keywords: Metacercariae; Opisthorchis felineus; Species-specific primers; Validation