title: Rapid and Sensitive Detection of Lymphocystis Disease Virus Genotype VII by Loop-Mediated Isothermal Amplification.
abstract: Lymphocystis disease virus (LCDV) infections have been described in gilthead seabream (Sparus aurata L.) and Senegalese sole (Solea senegalensis, Kaup), two of the most important marine fish species in the Mediterranean aquaculture. In this study, a rapid, specific, and sensitive detection method for LCDV genotype VII based on loop-mediated isothermal amplification (LAMP) was developed. The LAMP assay, performed using an apparatus with real-time amplification monitoring, was able to specifically detect LCDV genotype VII from clinically positive samples in less than 12 min. In addition, the assay allowed the detection of LCDV in all asymptomatic carrier fish analysed, identified by qPCR, showing an analytical sensitivity of ten copies of viral DNA per reaction. The LCDV LAMP assay has proven to be a promising diagnostic method that can be used easily in fish farms to detect the presence and spread of this iridovirus.
authors.affiliation: Departamento de Microbiología, Universidad de Málaga, 29071, Málaga, Spain.
authors.forename: Estefanía J
authors.initials: EJ
authors.lastname: Valverde
authors.affiliation: Aquatic Animal Disease, Centre for Environment, Fisheries and Aquaculture Science, The Nothe, Weymouth, Dorset, DT4 8UB, UK.
authors.forename: Irene
authors.initials: I
authors.lastname: Cano
authors.affiliation: Departamento de Microbiología, Universidad de Málaga, 29071, Málaga, Spain.
authors.forename: Dolores
authors.initials: D
authors.lastname: Castro
authors.affiliation: Aquatic Animal Disease, Centre for Environment, Fisheries and Aquaculture Science, The Nothe, Weymouth, Dorset, DT4 8UB, UK.
authors.forename: Richard K
authors.initials: RK
authors.lastname: Paley
authors.affiliation: Departamento de Microbiología, Universidad de Málaga, 29071, Málaga, Spain. jjborrego@uma.es.
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Borrego
mesh_terms: D000818:Animals; D017931:DNA Primers / Q000235:genetics; D004266:DNA Virus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D005393:Fish Diseases / Q000175:diagnosis / Q000821:virology*; D005838:Genotype; D007497:Iridoviridae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D021541:Sea Bream / Q000821:virology
chemical_list: D017931:DNA Primers
keywords: Carrier fish; Gilthead seabream; Lymphocystis disease virus; Real-time loop-mediated isothermal amplification; Senegalese sole