A　novel　bi-functionalized　aptasensor　for　thrombin　detection　was　fabricated　by　using　electrochemiluminescence　(ECL)　and　electrochemical　impedance　spectroscopy　(EIS)　techniques.　A　thiol-terminated　aptamer　with　15　oligonucleotides　was　hybridized　with　its　complementary　oligonucleotides　to　form　a　double-stranded　DNA　(dsDNA).　Then,　the　thiol-labeled　dsDNA　was　assembled　on　a　Au　electrode　via　Au-S　bond;　the　other　distal　of　the　dsDNA　labeled　with　biotin　was　bound　to　QDs　through　a　biotin-avidin　conjugation.　When　thrombin　is　present　in　the　detection　solution,　the　aptamer　can　combine　with　thrombin,　resulting　in　loss　of　CdSe　QDs　from　the　electrode　surface　and　thereby　decreasing　the　ECL　intensity　and　increasing　the　impedance.　The　decreased　ECL　and　increased　EIS　signals　are　logarithmically　linear　with　respect　to　the　concentration　Of　thrombin.　The　linear　range　was　10(-10)-10(-3)　mu　g　mL(-1)　(R=0.9924)　for　the　ECL　signal　and　10(-10)-10(-1)　mu　g　mL(-1)　(R=0.9875)　for　the　EIS　method　with　a　common　detection　limit　of　10(-10)　mu　g　mL(-1)　(2.7　aM).　In　addition,　the　bi-functionalized　aptasensor　exhibited　excellent　selectivity,　super　sensitivity,　a　low　detection　limit　and　a　wide　linear　range.　(C)　2015　Elsevier　B.V.　All　rights　reserved.