Articles
COMPARISON OF NON-RADIOACTIVE MOLECULAR HYBRIDIZATION METHODS AND RT-PCR FOR THE DETECTION OF HOP LATENT VIROID ON HOPS
Article number
656_30
Pages
187 – 191
Language
English
Abstract
Hop latent viroid (HLVd) is the serious hop plant (Humulus lupulus L.) pathogen which can be detected only by molecular biology methods.
Three methods for HLVd detection were compared.
The presence of the pathogen was examined by dot-blot hybridization and RT-PCR in the total nucleic acid extracts.
HLVd was also detected in fresh plant material by tissue print hybridization.
In this method the freshly cut petioles surfaces were pressed onto nylon membrane and after UV-fixing the membrane was subjected to hybridization.
DNA digoxigenin-labeled chemiluminescent detected probe was used both in dot-blot and tissue printing hybridization methods.
The results indicates that the both hybridization techniques can be useful in routine tests for HLVd detection and that negative signals obtained by these methods have to be checked by RT-PCR.
Three methods for HLVd detection were compared.
The presence of the pathogen was examined by dot-blot hybridization and RT-PCR in the total nucleic acid extracts.
HLVd was also detected in fresh plant material by tissue print hybridization.
In this method the freshly cut petioles surfaces were pressed onto nylon membrane and after UV-fixing the membrane was subjected to hybridization.
DNA digoxigenin-labeled chemiluminescent detected probe was used both in dot-blot and tissue printing hybridization methods.
The results indicates that the both hybridization techniques can be useful in routine tests for HLVd detection and that negative signals obtained by these methods have to be checked by RT-PCR.
Authors
M. Grudzińska, E. Solarska
Keywords
HLVd, detection, dot-blot, tissue print, RT-PCR
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