The Discrimination between Serotypes O and N of Potato virus Y Using a Multiplex PCR Assay

抄録

Enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies (Mabs) enabled to classify Potato virus Y (PVY) strains into two main serological groups: PVY^O,C serotype (O serotype) and PVY^N serotype (N serotype). This test is being used in seed potato certification programs and research laboratories involved in PVY characterization. However, Mabs are not always able to discriminate PVY strains due to the cross-reactivity of the Mabs, and the presence of recombination events in the capsid protein (CP) of some PVY isolates. Moreover, it is difficult to obtain these Mabs in some cases. In the present study, based upon specific nucleotide motifs in the CP gene of the serotypes O and N of PVY, we designed specific primers that can be used in multiplex PCR form for the simultaneous detection and identification of these serotypes. This assay can be performed during the molecular characterization of PVY isolates, hence eliminating the need for a separate ELISA test to identify PVY serotypes O and N. The reliability of this method was evaluated by comparing the results with the results of ELISA using well-documented Mabs for more than 80 PVY infected samples.