A diagnostic oligonucleotide microarray for simultaneous detection of grapevine viruses.

Area of Science:

• Plant Virology
• Molecular Diagnostics
• Agricultural Biotechnology

Background:

• Grapevine viruses cause significant global economic damage.
• Current detection methods (serological assays, RT-PCR) are time-consuming, especially for mixed infections.

Purpose of the Study:

• To develop and validate a 70-mer oligonucleotide microarray for simultaneous detection of grapevine viruses.
• To enable high-throughput plant certification and facilitate virus discovery.

Main Methods:

• Design of a microarray with 570 unique probes targeting 44 viral genomes and housekeeping genes.
• Utilized random primed RT-PCR amplification of double-stranded RNA from grapevine samples.
• Validated microarray accuracy against RT-PCR for 10 grapevine viruses and genomic libraries.

Main Results:

• The microarray successfully detected single and mixed grapevine virus infections.
• Consistent results were obtained when comparing microarray detection with RT-PCR.
• Three Closteroviridae members (GLRaV-4, -7, -9) were identified for the first time in Chilean grapevines.

Conclusions:

• The developed oligonucleotide microarray is a powerful tool for high-throughput grapevine virus detection and certification.
• This approach aids in the discovery of new viruses through cross-hybridization with conserved probes.
• The study identified novel viral infections in Chilean grapevines, highlighting the microarray's utility.