Clonagem de fragmentos genômicos de vírus quarentenários para o Brasil visando utilização como controle positivo em diagnose por PCR e RT-PCR

Abstract

The competitiveness generated by global agribusiness increases the Brazilian demand for plants capable of presenting higher productivity, without losing the quality of production. Therefore, Brazil needs to import vegetable material continuously, such as true seeds or other unit of vegetative propagation and seedlings. This enables to grow plants with a higher genetic potential, trhough genetic breeding or even by planting the imported material. However, it brings the imminent risk of introducing exotic pathogens into the country, such as quarentenary bacteria and virus, which are considered A1 Pest, potentially capable of causing great damages to Brazilian agriculture. Thus, the diagnosis of these pathogens in the imported material is crucial to prevent their introduction, which requires the use of reliable positive and negative controls, in order to avoid false positive and/or false negative. In this study genomic fragments of ten quarentenary viruses: African cassava mosaic virus (ACMV), Andean potato latent virus (APLV), Banana bract mosaic virus (BBrMV), Blueberry leaf mottle virus (BBLMV), Impatiens necrotic spot virus (INSV), Plum pox virus (PPV), Potato mop-top virus (PMTV), Potato yellowing virus (PYV), Potato virus T (PVT) and Tomato bushy stunt virus (TBSV), with potential to be used as positive controls in its diagnosis by RT-PCR and PCR, were cloned into a commercial plasmid. When tested, they behaved as good positive controls for the detection of the respective virus, using the primers designed specifically for this purpose. It is a safer alternative and avoids the need to use live pathogen, eliminating the risk of accidental introduction of exotic viruses in Brazil.