Novel strategy to evaluate infectious salmon anemia virus variants by high resolution melting.
Genetic variability is a key problem in the prevention and therapy of RNA-based virus infections. Infectious Salmon Anemia virus (ISAv) is an RNA virus which aggressively attacks salmon producing farms worldwide and in particular in Chile. Just as with most of the Orthomyxovirus, ISAv displays high variability in its genome which is reflected by a wider infection potential, thus hampering management and prevention of the disease. Although a number of widely validated detection procedures exist, in this case there is a need of a more complex approach to the characterization of virus variability. We have adapted a procedure of High Resolution Melting (HRM) as a fine-tuning technique to fully differentiate viral variants detected in Chile and projected to other infective variants reported elsewhere. Out of the eight viral coding segments, the technique was adapted using natural Chilean variants for two of them, namely segments 5 and 6, recognized as virulence-associated factors. Our work demonstrates the versatility of the technique as well as its superior resolution capacity compared with standard techniques currently in use as key diagnostic tools.
Laboratorio de Patógenos Acuícolas, Núcleo Biotecnología Curauma, Pontificia Universidad Católica de Valparaíso, Campus Curauma, Valparaíso, Chile.
Dagoberto
D
Sepúlveda
Constanza
C
Cárdenas
Marisela
M
Carmona
Sergio H
SH
Marshall
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D004279:DNA, Viral / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D016679:Genome, Viral; D008969:Molecular Sequence Data; D009975:Orthomyxoviridae / Q000145:classification* / Q000235:genetics; D016133:Polymerase Chain Reaction; D012474:Salmon / Q000821:virology*; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
D017931:DNA Primers; D004279:DNA, Viral