The　discrepancy　of　the　electrostatic　interaction　of　negatively　chargedsignal　molecules　to　long　and　short　DNA　strands　of　the　modified　electrode　surface　hasbeen　used　for　thefirst　time　to　the　develop　an　electrochemiluminescence　(ECL)biosensor　for　human　papillomavirus　16　(HPV　16)　DNA　detection.　The　short　single-stranded　capture　probe　(CP)　was　modifiedfirst　on　the　surface　of　the　gold　electrode,which　only　has　a　small　amount　of　negative　charge.　The　electrostatic　interactionbetween　the　negatively　charged　tris(2,2　＇-bipyridyl)　ruthenium(II)　chloride　hexahy-drate-doped　SiO2nanoparticles　(Ru@SiO2NPs)　and　CP　is　weak,　hence　Ru@SiO2NPs　easily　diffuse　to　the　surface　of　the　electrode　to　generate　a　strong　ECL　signal.Hybrid　chain　reaction　(HCR)　amplification　products　(long　strand　dsDNA)　wereprepared　in　homogeneous　solution　in　advance.　When　the　target　was　present,　the　dsDNA　can　be　connected　on　the　electrode　surfaceand　cause　the　enhancement　of　the　negative　charge　on　the　electrode　surface.　Owing　to　electrostatic　interaction　and　steric　hindrance,Ru@SiO2NPs　are　difficult　to　diffuse　to　the　electrode　surface,　resulting　in　a　significantly　reduced　ECL　signal.　The　decrease　of　ECLsignal　is　linearly　correlated　with　the　logarithm　of　the　HPV　concentration　under　optimal　conditions,　with　the　detection　range　being0.1　fM-5　pM　with　a　limit　of　1.41　aM.　This　innovative　methodology　expands　the　application　of　electrostatic　interaction　in　ECLsensing,　but　can　also　easily　develop　biosensors　for　detecting　other　targets　by　changing　the　DNA　sequence　used　in　this　strategy.