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Reverse transcription–detection of immobilized, amplified product in a one‐phase system (RT–DIAPOPS) for the detection of potato virus Y
Author(s) -
Nicolaisen M.,
Rasmussen H. N.,
Husted K.,
Nielsen S. L.
Publication year - 2001
Publication title -
plant pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.928
H-Index - 85
eISSN - 1365-3059
pISSN - 0032-0862
DOI - 10.1046/j.1365-3059.2001.00527.x
Subject(s) - biology , amplicon , reverse transcription polymerase chain reaction , potato virus y , reverse transcriptase , polymerase chain reaction , microbiology and biotechnology , virus , virology , chromatography , plant virus , gene , genetics , chemistry , gene expression
An RT–DIAPOPS (reverse transcription–detection of immobilized, amplified product in a one‐phase system) assay was developed for the detection of potato virus Y (PVY). The sensitivity of the assay was 0·5 pg when virions were added directly to the RT reaction. The assay was capable of detecting all isolates investigated, representing several taxonomic groups of PVY. The RT–DIAPOPS offers an attractive alternative to RT–polymerase chain reaction (PCR) as it does not depend on gel electrophoresis to detect virus‐specific PCR products. Finally, as amplicons are covalently attached to a solid phase, it minimizes the risk of contamination.