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Development and validation of real-time PCR assays based on novel molecular markers for the simultaneous detection and identification of Globodera pallida, G. rostochiensis and Heterodera schachtii

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Considering the growing trade of seed potato, reliable diagnostic protocols are required for the detection of regulated nematode species. In this study, a specific and sensitive multiplex Taqman-based real-time PCR method was developed in order to detect and identify Globodera pallida, G. rostochiensis and Heterodera schachtii. The newly designed primers and probes enabled the detection of all the target populations tested and with no cross-reaction for closely related non-target species (55 populations tested). The limit of detection (LOD) was one juvenile for G. rostochiensis and G. pallida and five juveniles for H. schachtii. For monitoring potato cyst nematodes, this analytical tool would extend the number of cyst investigated as five juveniles can be detected among 50 cysts in a sample. Furthermore, this multiplex assay detects DNA of the three targeted species in template DNA obtained directly from float material after nematode extraction from soil.

Affiliations: 1: 1Anses, Plant Health Laboratory, Nematology unit, Domaine de la Motte au Vicomte, BP35327, 35653 Le Rheu, France ; 2: 2INRA, UMR1349 IGEPP (Institute of Genetics, Environment and Plant Protection), Domaine de la Motte au Vicomte, BP35327, 35653 Le Rheu, France

*Corresponding author, e-mail:

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