Aspergillus　flavus　and　Aflatoxins　in　grain　crops　give　rise　to　a　serious　threat　to　food　security　and　cause　huge　economic　losses.　In　particular,　aflatoxin　B-1　has　been　identified　as　a　Class　I　carcinogen　to　humans　by　the　International　Agency　for　Research　on　Cancer　(IARC).　Compared　with　conventional　methods,　Surface-Enhanced　Raman　Scattering　(SERS)　has　paved　the　way　for　the　detection　of　Aspergillus　flavus　and　Aflatoxins　in　grain　crops　as　it　is　a　rapid,　nondestructive,　and　sensitive　analytical　method.　In　this　work,　the　rapid　detection　of　Aspergillus　flavus　and　quantification　of　Aflatoxin　B-1　in　grain　crops　were　performed　by　using　a　portable　Raman　spectrometer　combined　with　colloidal　Au　nanoparticles　(AuNPs).　With　the　increase　of　the　concentration　of　Aspergillus　flavus　spore　suspension　in　the　range　of　10(2)-10(8)　CFU/mL,　the　better　the　combination　of　Aspergillus　flavus　spores　and　AuNPs,　the　better　the　enhancement　effect　of　AuNPs　solution　on　the　Aspergillus　flavus.　A　series　of　different　concentrations　of　aflatoxin　B-1　methanol　solution　combined　with　AuNPs　were　determined　based　on　SERS　and　their　spectra　were　similar　to　that　of　solid　powder.　Moreover,　the　characteristic　peak　increased　gradually　with　the　increase　of　concentration　in　the　range　of　0.0005-0.01　mg/L　and　the　determination　limit　was　0.0005　mg/L,　which　was　verified　by　HPLC　in　ppM　concentration.　This　rapid　detection　method　can　greatly　shorten　the　detection　time　from　several　hours　or　even　tens　of　hours　to　a　few　minutes,　which　can　help　to　take　effective　measures　to　avoid　causing　large　economic　losses.