Detection ofGrapevine fanleaf virusby immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) with recombinant antibody

Grapevine fanleaf virus (GFLV), a member of the genus Nepovirus in the family Secoviridae, is the most widely distributed grapevine virus around the world. In the present research, recombinant antibody to the GFLV coat protein (CP) was used to develop an immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) for the detection of GFLV by the use of a pair of primers corresponding to the full length CP gene. In addition, double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was done by the use of recombinant antibody to detect the GFLV isolates. Eventually, an expected band with around 1512 base pair (bp) in length was revealed for three isolates by IC-RT-PCR. Also, DAS-ELISA revealed that the recombinant antibody was able to detect the virus in leaf sample. These results showed that application of recombinant antibody in these detection methods can be useful for rapid detection of viruses in plant tissue because by applying IC-RT-PCR along with recombinant antibody, it is possible to overcome the limitations happened by presence of inhibitors in grapevine trees and non-specific reaction resulting from presence of native proteins in plant sap against conventionally made antibody.