Certification protocol for the Grapevine Leafroll associated virus Type I (GLRaV-1)

Abstract

The 17,647 nucleotide genome sequence of GLRaV-1 has now been completed and sequence data analysed using bioinformatics. The shortcomings of the current tests for leafroll detection were dressed. The current RT-PCR test was inadequate due to poor reliability and reproducibility of results. The LRaV-1 RT-PCR certification protocol has been improved by using magnetic capture hybridisation prior to RT-PCR. The MCH-RT-PCR certification protocol has been demonstrated to provide a higher level of sensitivity and reproducibility than the current GLRaV-1 certification protocol used by the Australian industry. The MCH-RT-PCR technique provides an effective and practical screen to identify grapevine samples infected with GLRaV -1. The same protocol is applicable to other grapevine viruses.

Summary

The genome of Grapevine leafroll associated virus 1 (GLRaV-1) was fully characterized. Using the information, an improved GLRaV-1 certification protocol was developed by a magnetic capture-hybridization technique combined with PCR. The protocol was demonstrated to provide a higher level of sensitivity and reproducibility for the virus certification.