Detection method of potato spindle tuber viroid in true potato seeds with a non-radioactive DNA probe
A simple, sensitive and non-radioactive method for detecting potato spindle tuber viroid (EPPO A2 quarantine pest) in germinated true potato seeds has been developed. To produce seedlings, seeds are placed on moist filter paper in plastic boxes and allowed to germinate for 10 days in darkness at 25 °C. The method is based on nucleic acid hybridization with a PSTVd-specific DNA probe labelled with digoxigenin. The authors have found that with this method it was possible to detect one diseased seedling in approximately 150 healthy ones. This method can be used on ungerminated seeds but is less sensitive. They pointed out that this method is a good alternative to procedures based on phenol/chloroform extractions and hybridization procedures using radioactive probes and concluded that it could be of interested to most seed testing laboratories and institutions involved in the exchange of potato germplasm.
Borkhardt, B.; Vongsasitorn, D.; Albrechtsen, S.E. (1994) Chemiluminescent detection of potato spindle tuber viroid in true potato seed using a digoxigenin labelled DNA probe.
Potato Research, 37 (3), 249-255.