A diagnostic oligonucleotide microarray for simultaneous detection of grapevine virusesReport as inadecuate

A diagnostic oligonucleotide microarray for simultaneous detection of grapevine viruses - Download this document for free, or read online. Document in PDF available to download.


At least 58 viruses have been reported to infect grapevines causing economic damage globally. Conventionaldetection strategies based on serological assays, biological indexing and RT-PCR targeting one orfew viruses in each assay are widely used. Grapevines are prone to contain mixed infections of severalviruses, making the use of these techniques time-consuming. A 70-mer oligonucleotide microarray ableto detect simultaneously a broad spectrum of known viruses as well asnewviruses by cross-hybridizationto highly conserved probes is reported in the present study. The array contains 570 unique probesdesigned against highly conserved and species-specific regions of 44 plant viral genomes. In additionprobes designed against plant housekeeping genes are also included. By using a random primed RT-PCRamplification strategy of grapevine double stranded RNA-enriched samples, viral agents were detectedin single and mixed infections. The microarray accuracy to detect 10 grapevine viruses was comparedwith RT-PCR yielding consistent results. For this purpose, grapevine samples containing single or mixedinfections of Grapevine leafroll-associated virus-1 -2 -3 -4 -7 -9, Grapevine fanleaf virus, Grapevinerupestris stem pitting-associated virus, Grapevine virus A, and Grapevine virus B were used. Genomiclibraries containing complete viral genomes were also used as part of the validation process. The specificprobe hybridization pattern obtained from each virus makes this approach a powerful tool for highthroughput plant certification purposes and also for virus discovery if the new viral genomic sequenceshave partial similarity with the microarray probes. Three Closteroviridae members Grapevine leafrollassociatedvirus -4 -7 and -9 were detected for the first time in Chilean grapevines using the microarray.

Author: Engel, Esteban; - Escobar, Paula F.; - Rojas, Luis A.; - Rivera, Paulina; - Fiore, Nicola; - Valenzuela, Pablo D.T.; -

Source: http://repositorio.uchile.cl/


Related documents