A　highly　reproducible　ratiometric　aptasensor　has　been　developed　based　on　the　ratio　of　the　enhanced　electrochemiluminescence　(ECL)　signal　and　the　stable　electrochemistry　signal　for　the　first　time.　The　ECL　signal　tag　(Ru(phen)(3)(2+)-DNA)　was　chosen　as　working　probe　(Ru(phen)(3)(2+)+-wp)　and　electrochemistry　signal　tag　(methylene　blue-DNA)　as　reference　probe　(MB-rp).　The　hybridization　of　Ru(phen)(3)(2+)+-wp　and　MB-rp　in　one　structure　insured　the　interdependence　of　two　signals　on　the　same　sensing　interface.　OTA　was　chosen　as　a　target　model　in　this　work　and　substrate　DNA　(sDNA)　was　designed　to　hybridize　with　OTA-aptamer　(OTA-APT)　in　part,　as　well　as　the　triggered　chain　of　hybridization　chain　reaction　(HCR).　Only　ECL　signal　of　Ru(phen)(3)(2+)-wp　increased　with　the　OTA　existence　while　the　electrochemical　signal　from　MB-rp　was　stable　as　inter　reference　signal.　The　ratio　value　of　the　ECL/electrochemistry　had　a　linear　relationship　with　OTA　concentration　in　the　range　of　12.40　pM　to　6.19　nM　with　a　correlation　coefficient　of　0.990,　and　the　limit　of　detection　was　5.6　pM.　The　proposed　ECL　ratiometric　aptasensor　has　better　reproducibility　than　that　of　the　traditional　ECL　biosensor　and　been　applied　to　detect　OTA　in　real　samples　with　satisfied　results.　(C)　2018　Elsevier　Ltd.　All　rights　reserved.