A diagnostic oligonucleotide microarray for simultaneous detection of grapevine virusesReportar como inadecuado




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Resumen

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.



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

Fuente: http://repositorio.uchile.cl/



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