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Real-time PCR for the detection and quantitative analysis of IHNV in salmonids
Authors:K Overturf,S LaPatra,&   M Powell
Affiliation:USDA-Agricultural Research Service (ARS), Hagerman, ID, USA;Clear Springs Foods, Inc, Research Division, Buhl, ID, USA;University of Idaho, Hagerman Fish Culture Experiment Station, Hagerman, ID, USA
Abstract:The rapid identification and quantification of virus in diseased fish is a goal both conservationists and commercial aquaculturists have struggled to attain. Recently a technique for the detection of viral mRNA particles that uses fluorescent tagging and amplification has been developed. Utilizing primers and fluorescent labelled probes generated for the specific identification of the nucleocapsid (N) and glycoprotein (G) genes of infectious haematopoietic necrosis virus (IHNV), and an instrument that measures cyclic emittance of fluorescence, the presence or absence of virus can be easily and rapidly confirmed. This method is not only useful in confirming viral presence but is effective in measuring the relative or absolute quantity of virus present within the sample. This allows for the determination of the health status of a carrier fish by measuring the quantity of viral genomes or transcribed viral genes present. Because this method is based on sequence detection, instead of virus isolation in cell culture, it is also effective in determining the presence of pathogenic organisms from water, fish feeds, or other potential reservoirs of infection.
Keywords:IHNV    detection    quantification    PCR
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