Sulfonamide　antibiotics　(SAs)　are　widely　used　in　animal　husbandry　and　aquaculture,　and　the　excess　residues　of　SAs　in　animal-derived　foods　will　harm　the　health　of　consumers.　In　reality,　various　SAs　were　alternately　used　in　animal　husbandry　and　aquaculture,　and　thus,　it　is　urgent　need　to　develop　simple　and　high-throughput　methods　for　simultaneously　detecting　multiple　SAs　or　groups　of　SAs　in　order　to　realize　rapid　screening　of　total　SAs　residues　in　animal-derived　foods.　We　herein　isolated　a　broad-specificity　aptamer　for　SAs　by　using　a　multi-SAs　systematic　evolution　of　ligands　by　exponential　enrichment　(SELEX)　strategy.　The　isolated　broad-specificity　aptamer　has　a　higher　binding　affinity　to　five　different　SAs　including　sulfaquinoxaline　(SQ),　sulfamethoxypyridazine　(SMPZ),　sulfametoxydiazine　(SMD),　sulfachloropyridazine　(SCP),　and　sulfapyridine　(SPD)　and,　thus,　can　be　used　as　a　bioreceptor　for　developing　various　high-throughput　methods　for　the　simultaneous　detection　or　rapid　screening　of　above　five　SAs.　Based　on　the　isolated　broad-specificity　aptamer　and　Cy7　(diethylthiatricarbocyanine)　displacement　strategy,　a　colorimetric　aptasensor　was　developed　for　the　simultaneous　detection　of　SQ,　SMPZ,　SMD,　SCP,　and　SPD　with　a　visual　detection　limit　of　2.0-5.0　μM　and　a　spectrometry　detection　limit　of　0.2-0.5　μM.　The　colorimetric　aptasensor　was　successfully　used　to　detect　SQ,　SMPZ,　SMD,　SCP,　and　SPD　in　fish　muscle　with　a　recovery　of　82%-92%　and　a　RSD　(n　=　5)　©　2022　American　Chemical　Society.　All　rights　reserved.