Ahighlysensitiveelectrochemiluminescence-polymerasechainreaction(ECL-PCR)methodforK-raspointmutationdetectionisdeveloped.Briefly,K-rasoncogenewasamplifiedbyaRu(bpy)32+(TBR)-labeledforwardandabiotin-labeledreverseprimer,andfollowedbydigestionwithMvaIrestrictionenzyme,whichonlycutthewild-typeampliconcontainingitscuttingsite.Thedigestedproductwasthenadsorbedtothestreptavidin-coatedmicrobeadthroughthebiotinlabelanddetectedbyECLassay.TheexperimentresultsshowedthatthedifferentgenotypescanbeclearlydiscriminatedbyECL-PCRmethod.Itisusefulinpointmutationdetection,duetoitssensitivity,safety,andsimplicity.
