Validation of quantitative DNA detection systems for PCN

Traditional methods to detect and identify PCN in soil involve separating cysts from soil, for example using a Fenwick can or Wye washer, followed by identification of the cysts either using a microscope or DNA-based methods.

A technique (TaqMan assay) that detects the presence of PCN DNA was developed and used in conjunction with extracted cysts. Methods for the direct extraction of DNA from soil samples were also tested in conjunction with the TaqMan assay. Methods to extract DNA from volumes of soil up 1.5kg were developed and shown to work.

Overall, the methods developed can be tailored to specific needs yielding cyst numbers, egg numbers and individual species from between 250g and 1.5kg soil depending on specific requirements. The costs vary according to the level of information required and volume of soil processed.

To develop rapid, DNA-based diagnostics to quantify the amount of Globodera pallida and/or G. rostochiensis in a soil sample.