Imidacloprid, acephate, and carbaryl are common insecticides that are extensively used in planting mango, a well-known fruit in Vietnam, to ease mango hopper issues. The accurate detection of pesticide residues is critical for mango export to meet quality criteria. This study developed a novel SERS platform by using polydimethylsiloxane (PDMS) to simulate the rose petal structure incorporated with a silver coating layer and silver nanoparticles (AgNPs) to detect imidacloprid, acephate, and carbaryl in mango fruits. In this paper, the rose petal PDMS/Ag-AgNPs replica was considered the most efficient substrate for SERS measurement with an EF of 4.7 × 107. The Raman spectra of the three insecticides obtained from the PDMS/Ag-AgNPs substrate were clearly observed with their characteristic peaks of 1105 cm−1 for imidacloprid, 1083 cm−1, and 1579 cm−1 for acephate, and 727 cm−1 and 1378 cm−1 for carbaryl. The application of PDMS/Ag-AgNPs substrate in quantitative analysis of the three pesticides in mango fruit was evaluated. As a result, the limit of detection was 0.02 mg/kg for imidacloprid, 5 × 10−5 mg/kg for acephate, and 5 × 10−3 mg/kg for carbaryl. The SERS result also revealed that the pesticide residues in the mango sample were within an acceptable limit. This suggested the possibility of the rose petal PDMS/Ag-AgNPs replica for rapid quantification of pesticide residues not only in mango fruit but also in many other agricultural products.