In　this　work,　we　have　developed　a　double-layer　nanofiber　membrane　(NFM)　with　high　selectivity　based　on　molecular　imprinting　technique　by　electrospinning　methods　and　applied　it　in　recognizing　putrescine.　Poly(vinyl　alcohol)　(PVA)　was　used　as　spinning　solution,　1,4-butylene　glycol　as　the　imitate　template,　and　ninhydrin　as　chromogenic　agent.　The　adsorption　of　the　putrescine　is　corresponded　to　purple-color　stains　on　the　NFM,　which　is　digitizing　intuitively　by　the　ImageJ　software.　By　quantifying　the　gray　value　of　the　stains　on　the　NFM,　its　imprinting　efficiency　(IE)　is　estimated.　The　adsorption　performance　of　the　double-layer　molecular　imprinted　NFM　was　studied　in　detail,　showing　higher　adsorption　capacity　(71.912　mg　mL(-1)),　shorter　adsorption　equilibrium　time　of　40　min　and　higher　selectivity　to　putrescine　than　our　previous　works.　Furthermore,　the　material　showed　remarkable　selectivity　for　putrescine　with　respect　to　other　three　kinds　of　aliphatic　amine　in　similar　molecular　structure.　The　selectivity　coefficient　(alpha)　of　putrescine　against　cadaverine,　spermine,　and　spermidine　are　calculated　to　be　1.734,　2.224,　and　2.576,　respectively.　In　addition,　since　PVA　is　a　water-soluble　material,　the　NFM　is　not　reusable.　(c)　2020　Wiley　Periodicals,　Inc.　J.　Appl.　Polym.　Sci.　2020,　137,　48932.