Direct tuber testing for Potato Y potyvirus by real-time RT-PCR and ELISA: reliable options for post-harvest testing?*


  • *

    Paper presented at the EPPO Conference on Quality of Diagnosis and New Diagnostic Methods for Plant Pests (Noordwijkerhout, NL, 2004-04-19/22).


The method currently used for testing potato tubers for viruses following harvest involves a growing-on test. This takes up to 6 weeks to complete, and there is therefore a demand for more rapid test results. The sensitivity and reliability of direct tuber testing by DAS-ELISA and real-time RT-PCR (TaqMan) were compared with the growing-on test. In addition, the reliability of all three methods for the detection of Potato Y potyvirus (PVY) in tubers was compared over post-harvest intervals of 6, 10, 14 and 18 weeks. The test material came from plots of tubers (cv. ‘Maris Piper’) containing a primary infection of strains PVYN and PVYO, following aphid transmission from marked infector plants grown during the 2003 season. Sample material was homogenized and divided, to provide comparative test material for detection of PVY by ELISA and real-time RT-PCR. Tuber eye-plugs were then taken and subjected to the growing-on test. The remainder of the tuber was also grown on and tested, to ensure infection was not missed as a consequence of an uneven distribution of virus throughout the tuber material. The results obtained using the two methods for direct testing of the tubers, and those results obtained from the traditional growing-on test, are compared. The advantages and disadvantages of each method are discussed.