Development of RPA-EXO assay for detection of potato golden nematode Globodera rostochiensis
Potato cyst nematode(PCN)is an important quarantine pest in the world and causes major loss in potato crops.In this study,RPA(Recombinase polymerase amplification)specific primers and flu-orescent probe were designed based on the ITS sequence of Globodera rostochiensis,and an RPA-EXO detection method for G.rostochiensis was established.By optimizing the reaction time and temperature of RPA,the optimal conditions for RPA detection method were 38℃ for 30 min.This assay can distinguish G.rostochiensis from five other plant nematode populations.The sensitivity of RPA-EXO detection method for G.rostochiensis can be achieved 10-3 single cyst DNA.This is the method for diagnosis G.ros-tochiensis using RPA-EXO,it's a rapid,convenient and sensitive detection assay.It provides a new rapid method for the detection of G.rostochiensis for grassroots personnel.
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