Protocols

In order to achieve international harmonisation of the testing protocol for the detection of pospiviroids in tomato and sweet pepper seeds, Naktuinbouw makes its protocols available to other parties.

These protocols can be used for detecting CEVd, CLVd, MPVd, PCFVd, PSTVd, TASVd, TCDVd and TPMVd. The protocols were developed in a 3x1000 seed format and are also available in a 50x400 seed format to meet Australian requirements. They involve manual RNA extraction (for sweet pepper seeds) or KingFisher extraction (for tomato seeds). The RT-PCR is based on a combination of a number of multiplex PCRs for detecting eight pospiviroids. The detection of PSTVd-TCVd is based on the current Naktuinbouw protocol for these viroids, which has been validated. The entire protocol will be validated in the EU-TESTA project.

Detection of pospiviroid in tomato

Detection of pospiviroid in pepper

Compliance ISHI method