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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
immunology and microbiology
Oligonucleotide microarray-based detection and genotyping of Plum pox virus
Journal of Virological Methods, Volume 147, No. 1, Year 2008
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Description
Plum pox virus (PPV) is the most damaging viral pathogen of stone fruits. The detection and identification of its strains are therefore of critical importance to plant quarantine and certification programs. Existing methods to screen strains of PPV suffer from significant limitations such as the simultaneous detection and genotyping of several strains of PPV in samples infected with different isolates of the virus. A genomic strategy for PPV screening based on the viral nucleotide sequence was developed to enable the detection and genotyping of the virus from infected plant tissue or biological samples. The basis of this approach is a long 70-mer oligonucleotide DNA microarray capable of simultaneously detecting and genotyping PPV strains. Several 70-mer oligonucleotide probes were specific for the detection and genotyping of individual PPV isolates to their strains. Other probes were specific for the detection and identification of two or three PPV strains. One probe (universal), derived from the genome highly conserved 3′ non-translated region, detected all individual strains of PPV. This universal PPV probe, combined with probes specific for each known strain, could be used for new PPV strain discovery. Finally, indirect fluorescent labeling of cDNA with cyanine after cDNA synthesis enhanced the sensitivity of the virus detection without the use of the PCR amplification step. The PPV microarray detected and identified efficiently the PPV strains in PPV-infected peach, apricot and Nicotiana benthamiana leaves. This PPV detection method is versatile, and enables the simultaneous detection of plant pathogens. © 2007 Elsevier B.V. All rights reserved.
Authors & Co-Authors
Pasquini, Graziella
Italy, Rome
Istituto Sperimentale Per la Patologia Vegetale
Barba, Marina
Italy, Rome
Istituto Sperimentale Per la Patologia Vegetale
Hadidi, Ahmed F.
United States, Beltsville
Usda Ars Beltsville Agricultural Research Center
Faggioli, F.
Italy, Rome
Istituto Sperimentale Per la Patologia Vegetale
Negri, Rodolfo
Italy, Rome
Sapienza Università Di Roma
Sobol, Iris
Israel, Jerusalem
Hebrew University of Jerusalem
Tiberini, Antonio
Italy, Rome
Istituto Sperimentale Per la Patologia Vegetale
Çaǧlayan, Kadriye
Turkey, Antakya
Mustafa Kemal Üniversitesi
Mazyad, Hamed M.
Egypt, Cairo
Egyptian Ministry of Agriculture, Cairo
Anfoka, Ghandi H.
Jordan, Al Salt
Al-balqa Applied University
Ghanim, Murad
Israel, Bet Dagan
Agricultural Research Organization of Israel
Zeidan, Mohammad
Israel, Bet Dagan
Israel Ministry of Agriculture and Rural Development
Czosnek, Henryk H.
Israel, Jerusalem
Hebrew University of Jerusalem
Statistics
Citations: 47
Authors: 13
Affiliations: 9
Identifiers
Doi:
10.1016/j.jviromet.2007.08.019
ISSN:
01660934
Research Areas
Genetics And Genomics