Development and Validation of a Loop-Mediated Isothermal Amplification Diagnostic Method to Detect the Quarantine Potato Pale Cyst Nematode, Globodera pallida.

Eugénia De Andrade,Manuel Mota,Maria L Inácio,Maria João Camacho

doi:10.3390/pathogens10060744

Eugénia De Andrade, Manuel Mota + Show 2 more

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https://doi.org/10.3390/pathogens10060744

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Abstract

The potato cyst nematode (PCN) Globodera pallida 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 G. pallida 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 G. pallida in a sample of at least 40 juveniles of the non-target species G. rostochiensis. This test should be a tool to improve the performance of the laboratory and has the potential to be performed on-site.

Highlights

IntroductionThe potato cyst nematodes (PCN), Globodera rostochiensis [1,2] and Globodera pallida [3]
The objective of this work is to develop a loop-mediated isothermal amplification method (LAMP) assay for G. pallida, to be used in routine analyses, since the analysis of soils sampled in Portuguese potato fields has revealed an increased spread of G. pallida in the country [52]
Over, costs associated with the damage caused by new pests in the invaded areas as a costs associated with the damage caused by new pests in the invaded areas as a result of result of decreases in production, market and pest management, surveillance and decreases in production, market value andvalue pest management, surveillance and inspection inspection may benefit from an early detection

Summary

Introduction

The potato cyst nematodes (PCN), Globodera rostochiensis [1,2] and Globodera pallida [3]. Constitute one of the greatest threats to potato crops. These plant parasitic nematodes originated from the Andes region in southern Peru and have spread as the result of anthropogenic activity into many regions of the world [4]. They are thought to have been introduced into Europe in the 16–17th century by means of potato tubers carrying infested soil. Beyond Europe, PCN have been reported throughout South America and parts of

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