In this study, we used a starch paste stabilizer to synthesize ZnSe: Mn/ZnS- Starch and ZnSe/ZnS: Mn/ZnS-starch quantum dot (QDs) in a non-toxic aqueous solvent. The -CH2-OH group of the starch paste promotes dispersibility and improves the compatibility of quantum dots with antibodies, its bonding is observed in the FTIR spectrum. Besides, the Mn-doped ZnS buffer shell with various concentrations (1, 3, 5, 7, and 9%) influence structure, optical, and photoluminescence of QDs properties were investigated in detail. The greatest luminescence intensity is achieved at a molar ratio of 3% Mn2+/Zn2+. Moreover, the ZnS: Mn buffer shell helps to enhance the fluorescence intensity and quantum yield (QY) of the ZnSe/ZnS: Mn/ZnS QDs, which are higher than ZnSe: Mn/ZnS-starch QDs. Through protein A and EDC bridging, ZnSe/ZnS:3%Mn/ZnS- Starch resulted in good signal and sensitivity, with no toxicity to E. coli O157:H7 and MRSA strains.