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A detection method for infectious pancreatic necrosis virus (IPNV) based on reverse transcription (RT)‐polymerase chain reaction (PCR)

Authordc.contributor.authorLópez Lastra, Marcelo 
Authordc.contributor.authorGonzález Canales, Marcelo 
Authordc.contributor.authorJashes, M. 
Authordc.contributor.authorSandino García, Ana María 
Cita de ítemdc.identifier.citationJournal of Fish Diseases, Volumen 17, Issue 3, 1994, Pages 269-282
Abstractdc.description.abstractA rapid, sensitive and highly specific detection method for infectious pancreatic neerosis virus (IPNV), based on reverse transcription (RT) polymerase chain reaction (PCR) has been developed. The specificity of the assay is provided by the oligonucleotide primers selected from the IPNV major capsid polypeptide VP2 gene. For each primer combination only one major product is obtained when amplifieation is performed using IPNV double‐stranded RNA from two different viral strains, Sp and VR‐299, as the initial template. No products were detected when genomie nueleic acids other than IPNV RNA were used as RT‐PCR templates. The specificity of the amplification products were confirmed by Southern hybridization using a specific cDNA probe. To assess the sensitivity of the method, dilutions of purified IPNV dsRNA total genome were amplified and quantities of as little as 1 pg of purified dsRNA were detected when the amplification product was visualized by silver‐stained polyacrylamidc gel electrophoresis. This technique detected IPNV directly in infected coho salmon, Oncorhynchus kisutch (Walbaum), and rainbow trout, Oncorhynchus mykiss (Walbaum), tissues and fish egg samples, avoiding viral propagation in cell culture. The results show that this RT‐PCR amplification method is useful for the direct tissue detection of IPNV
Sourcedc.sourceJournal of Fish Diseases
Keywordsdc.subjectAquatic science
Keywordsdc.subjectVeterinary (miscellaneous)
Títulodc.titleA detection method for infectious pancreatic necrosis virus (IPNV) based on reverse transcription (RT)‐polymerase chain reaction (PCR)
Document typedc.typeArtículo de revista
Indexationuchile.indexArtículo de publicación SCOPUS
Access notedct.AccessRightsSin acceso a texto completo

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