title: High-throughput diagnosis of potato cyst nematodes in soil samples.
abstract: Potato cyst nematode (PCN) is a damaging soilborne pest of potatoes which can cause major crop losses. In 2010, a new European Union directive (2007/33/EC) on the control of PCN came into force. Under the new directive, seed potatoes can only be planted on land which has been found to be free from PCN infestation following an official soil test. A major consequence of the new directive was the introduction of a new harmonized soil sampling rate resulting in a threefold increase in the number of samples requiring testing. To manage this increase with the same staffing resources, we have replaced the traditional diagnostic methods. A system has been developed for the processing of soil samples, extraction of DNA from float material, and detection of PCN by high-throughput real-time PCR. Approximately 17,000 samples are analyzed each year using this method. This chapter describes the high-throughput processes for the production of float material from soil samples, DNA extraction from the entire float, and subsequent detection and identification of PCN within these samples.
authors.affiliation: Science and Advice for Scottish Agriculture, Roddinglaw Road, Edinburgh, EH12 9FJ, UK, alex.reid@sasa.gsi.gov.uk.
authors.forename: Alex
authors.initials: A
authors.lastname: Reid
authors.forename: Fiona
authors.initials: F
authors.lastname: Evans
authors.forename: Vincent
authors.initials: V
authors.lastname: Mulholland
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Cole
authors.forename: Jon
authors.initials: J
authors.lastname: Pickup
mesh_terms: D000818:Animals; D018509:DNA, Helminth / Q000032:analysis* / Q000235:genetics; D018744:DNA, Plant / Q000032:analysis* / Q000235:genetics; D009348:Nematoda / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification* / Q000472:pathogenicity; D010572:Pest Control, Biological / Q000379:methods*; D010935:Plant Diseases / Q000469:parasitology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012987:Soil / Q000469:parasitology*; D011198:Solanum tuberosum / Q000469:parasitology*
chemical_list: D018509:DNA, Helminth; D018744:DNA, Plant; D012987:Soil