| Summary: | The potato cyst nematode (PCN) <i>Globodera pallida</i> has acquired significant importance throughout Europe due to its nefarious effects on potato production. Rapid and reliable diagnosis of PCN is critical during the surveillance programs and for the implementation of control measures. Molecular DNA-based methods are available, but they require expensive laboratory facilities, equipment and trained technicians. Moreover, there is an additional need of time for sample shipment and testing. In this work, we have developed a new and simple assay which reliably discriminates <i>G. pallida</i> from other cyst nematodes in less than 40 min. This assay may be applied either on cysts or juveniles with the ability to detect a single juvenile of <i>G. pallida</i> in a sample of at least 40 juveniles of the non-target species <i>G. rostochiensis</i>. This test should be a tool to improve the performance of the laboratory and has the potential to be performed on-site.
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