摘要： Bio-functionalization of quantum dots (QDs) is of important value in practical applications. With single-stranded DNA (ssDNA) rich in thymine T and thioguanine G taken as the template, a new-type nanocomposite material (ssDNA-PQDs) synthesized from low-toxicity T-ssDNA functionalized Mn–ZnS and room-temperature phosphorescent (RTP) QDs (PQDs) was prepared in this paper by optimizing synthesis conditions, and these ssDNA-PQDs could emit orange RTP signals at 590 nm. As these ssDNA-PQDs are rich in T sequences and T sequences can bond with melamine through the hydrogen-bond interaction, ssDNA-PQDs experience aggregation, thus causing phosphorescent exciton energy transfer (PEET) between ssDNA-PQDs of di?erent particle sizes and their RTP quenching. Based on this principle, an RTP detection method for melamine was established. The linear range and detection limit of the detection method are 0.005–6 mM and 0.0016 mM respectively. As this method is based on the RTP nature of ssDNA-PQDs, it can avoid disturbance from background ?uorescence and scattered light of the biological ?uid, and it is very suitable for melamine detection in the biological ?uid milk.