Report Open Access
Kiewnick, Sebastian; Braun-Kiewnick, Andrea; Viaene, Nicole; Hallmann, Johannes; Niere, Bkörn; Gamel, Sylvie; Roversi, Pio Federico; den Nijs, Loes; van de Vossenberg, Bart; Toktay, Halil; Prior, Tom; Sapp, Melanie
The aim of this Euphresco project was to coordinate ongoing and future research in the field of Meloidogyne enterolobii detection and identification and to validate new methods in a collaborative way. It contained six main topics:
- Test performance study on nematode extraction methods to detect motile stages (second stage juveniles) of M. enterolobii in soil samples.
- Test performance study on fast and simple DNA extraction from target nematode in complex nematode suspension after extraction from soil samples (Lysis buffer; Holterman et al., 2009). DNA needs to be clean enough (without inhibitors) for further by molecular detection/identification tools. Labs were encouraged but not required to use additional extraction methods. The aim was to test a fast and simple DNA extraction protocol that can be harmonized between NPPO laboratories in the future.
- Methods development and in-house validation of qPCR methods to detect, identify and potentially quantify M. enterolobii in complex nematode suspensions obtained from soil samples (develop protocols for robust, specific and sensitive detection/identification that can be harmonized between NPPO labs, independent of lab equipment and chemistry).
- Test performance study on protocols developed under topic 3.
- Final meeting to communicate and discuss results. Optional workshop on Barcoding as new diagnostic tool for fast and reliable nematode identification was offered to interested groups.
- Publication of results in peer-reviewed research journals and discussion for inclusion of protocols into EPPO diagnostic standard protocol. Identification of future research priorities in the field of Meloidogyne enterolobii research.