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Comparative analysis of ELISA, nonradioactive molecular hybridization and PCR for the detection of prunus necrotic ringspot virus in herbaceous and Prunus hosts
Author(s) -
J. A. SánchezNavarro,
Frederic Aparicio,
Adib Rowhani,
Vicente Pallás
Publication year - 1998
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.1365-3059.1998.00301.x
Subject(s) - biology , prunus , polymerase chain reaction , microbiology and biotechnology , herbaceous plant , virus , virology , dot blot , detection limit , gene , botany , chromatography , chemistry , genetics
Three methods were compared for the detection of prunus necrotic ringspot virus in herbaceous and woody plants: DAS‐ELISA, nonisotopic dot‐blot hybridization and reverse transcriptional polymerase chain reaction (RT‐PCR). When purified virus preparations were used, the detection limit of the RT‐PCR technique was 1.28 pg mL −1 whereas nonisotopic molecular hybridization and DAS‐ELISA allowed detection of 0.8 ng mL −1 and 4 ng mL −1 , respectively. Several sample processing procedures were evaluated for virus detection by the nonisotopic molecular hybridization technique. When a very short and simple sample processing method was used, the detection limit of the nonisotopic molecular hybridization technique was 25 times higher than that of DAS‐ELISA and 625 times lower than that of RT‐PCR. A comparison of the level of virus accumulation in mature fruits and in leaf tissue showed that, on average, 125 times more virus was found in the fruits.