Development of a loop-mediated isothermal amplification (LAMP) assay for the identification of the invasive wood borer Aromia bungii (Coleoptera: Cerambycidae) from frass

The red-necked longhorn beetle Aromia bungii (Faldermann, 1835) (Coleoptera: Cerambycidae) is native to east Asia, wherit is a major pest of cultivated and ornamental species of the genus Prunus. Morphological or molecular discrimination ofadults or larval specimens is required to identify this invasive wood borer. However, recovering larval stages of the pest fromtrunks and branches causes extensive damage to plants and is timewasting. An alternative approach consists in applying noninvasivemolecular diagnostictoolstobiological traces(i.e., fecalpellets,frass).In thisway,infestationsin hostplants canbedetectedwithoutdestructivemethods.Thispaperpresentsaprotocolbasedonbothreal-timeandvisualloop-mediatedisothermalamplification(LAMP),using DNAof A.bungiiextractedfromfecalparticlesin larvalfrass.Laboratoryvalidationsdemonstratedtherobustnessof theprotocolsadoptedand theirreliabilitywasconfirmedperformingan inter-labblindpanel.The LAMPassayand theqPCR SYBRGreenmethodusing theF3/B3 LAMPexternalprimerswereequallysensitive,andbothweremoresensitivethantheconventionalPCR (sensitivity>10 to the same starting matrix). The visual LAMPprotocol, due to the relatively easy performance of the method, could be a useful tool to apply in rapid monitoring of Abungii and in the management of its outbreaks.3