title: Denaturing gradient gel electrophoresis (DGGE) as a powerful novel alternative for differentiation of epizootic ISA virus variants.
abstract: Infectious Salmon Anemia is a devastating disease critically affecting world-wide salmon production. Chile has been particularly stricken by this disease which in all cases has been directly related with its causative agent, a novel orthomyxovirus which presents specific and distinctive infective features. Among these, two molecular markers have been directly associated with pathogenicity in two of the eight RNA sub genomic coding units of the virus: an insertion hot spot region present in viral segment 5 and a Highly Polymorphic Region (HPR) located in viral segment 6. Here we report the successful adaptation of a PCR-dependent denaturing gel electrophoresis technique (DGGE), which enables differentiation of selected reported HPR epizootic variants detected in Chile. At the same time, the technique allows us to distinguish one nucleotide differences in sequences associated with the intriguing, and still not well-understood, insertion events which tend to occur on RNA Segment 5. Thus, the versatility of the technique opens new opportunities for improved understanding of the complex biology of all ISA variants as well as possible applications to other highly variable pathogens.
authors.affiliation: Laboratorio de Patologenos Acuícolas, Núcleo Biotecnología Curauma, Pontificia Universidad Católica de Valparaíso, Campus Curauma, Valparaíso, Chile.
authors.forename: Marisela
authors.initials: M
authors.lastname: Carmona
authors.forename: Dagoberto
authors.initials: D
authors.lastname: Sepúlveda
authors.forename: Constanza
authors.initials: C
authors.lastname: Cárdenas
authors.forename: Luis
authors.initials: L
authors.lastname: Nilo
authors.forename: Sergio H
authors.initials: SH
authors.lastname: Marshall
mesh_terms: D000818:Animals; D002677:Chile; D017931:DNA Primers / Q000235:genetics; D058645:Denaturing Gradient Gel Electrophoresis / Q000379:methods*; D005393:Fish Diseases / Q000175:diagnosis* / Q000821:virology*; D046829:Isavirus / Q000235:genetics* / Q000302:isolation & purification*; D009976:Orthomyxoviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D011110:Polymorphism, Genetic / Q000235:genetics; D020317:Salmo salar; D013045:Species Specificity
chemical_list: D017931:DNA Primers