Effect of Salmonid alphavirus infection on housekeeping gene expression, validation by quantitative, real-time PCR

Abstract

Pancreas disease is responsible for major economic losses in the European salmonid farming industry, due to mortality, diminished growth rate and reduced slaughter weights. The infectious agent causing pancreas disease in Atlantic salmon was first isolated in Scotland in 1995 and later classified as an Alphavirus, the first Alphavirus to be isolated that affects fish. One hallmark of Alphavirus infection in vertebrate cells is the ability of the pathogen to shut down host transcription and translation processes without affecting viral protein and nucleic acid synthesis. It is of interest to find out if SAV shares this characteristic. The goal of this project was to investigate Salmonid alphaviruses ability to down-regulate host cell transcription. Through using a two-step quantitative PCR for analysis of different housekeeping genes in non-infected and infected cells, we hoped to evaluate if SAV3 causes down-regulation of transcription. If so, at what time interval it occurred, and to what degree. This study and its incurred results served as a pilot study, to optimize and fine-tune the methodology for further experimentation.