Comparison of rapid detection assays for grapevine leafroll disease associated closteroviruses

Three rapid detection assays (ELISA, dsRNA analysis and ISEM) were compared for their sensitivity, specificity, and simplicity in the detection of grapevine leafroll associated closteroviruses (GLRaV). Each was found to have advantages and disadvantages for routine testing. ELISA is sensitive and easy to use, but different antisera are needed to detect different GLRaV types. Because mixing or blending of antisera can produce good results in a single ELISA test, each antiserum does not need to be used separately unless it is important to determine the type of GLRaV present. DsRNA analysis can detect all the types of GLRaV tested but has a relatively low sensitivity and is labor intensive, which makes it unsuitable for testing large numbers of samples. Furthermore, dsRNA does not give unequivocal diagnosis of GLRaV infections. ISEM is sensitive and rapid. However, like ELISA, this technique requires an antiserum to each GLRaV type tested and an electron microscopy. Our recommendation is that ELISA should be used with multiple antisera for large scale testing programs. Samples for which ELISA results are inconclusive should be retested with ISEM and/or dsRNA. When the disease status of an individual sample must be determined conclusively, a few grams of tissue should be processed to concentrate the virus and then subjected to ELISA and examination by electron microscopy with negative staining. A dsRNA analysis should be carried out as well.