DTBA and ELISA methods in detection of grapevine leafroll-1 virus

Abstract

Grapevine leafroll syndrome is caused by a complex of up to nine different Grapevine leafroll-associated viruses (GLRaV-1-9). Many methods, including indexing, serological and molecular procedures, have been developed for the detection of GLRaV. However, due to the low concentration of the virus in plants a method with improved sensitivity and with the capacity to detect is required. In this study plants were tested for Grapevine leafroll virus (GLRaV) by visual examination, enzyme-linked immunosorbent assay (ELISA) and direct tissue-blotting assay (DTBA). Tissues were homogenized for use in a double antibody sandwich ELISA system and then blotted on a nitrocellulose membrane for DTBA. Comparison of DTBA with ELISA and with visual plant symptoms suggest that DTBA can be used with the same accuracy as ELISA for detecting GLRaV-1 in leaf tissue.

Sindrom uvijanja lista vinove loze je oboljenje koje prouzrokuje grupa virusa koja se naziva Virusi vezani za uvijanje lista vinove loze (GLRaV-1-9). Više različitih metoda je razvijeno za detekciju ove grupe virusa, kao što su indeksiranje, serološke i molekularno biološke metode. Zbog relativno male koncentracije virusa u zaraženoj biljci potrebno je pronaći metodu koja ima najveću osetljivost pri njegovoj detekciji. U ovom radu u cilju detektovanja virusa GLRaV, ispitivane biljke su pored vizuelnog pregleda obrađivane i pomoću ELISA i DTBA metoda. Poređenjem ELISA i DTBA metode na simptomatičnim biljkama utvrđeno je da se ove dve metode sa jednakom tačnošću mogu koristiti u otkrivanju GLRaV-1 u listu vinove loze.