In this report, polylactide (PLA) nanofibers were fabricated with the new strategy of electrospinning that may have some potential prominent application in biomedical areas. Besides, the new nanocomposites produced by the combination of PLA nanofibers with titanium dioxide (TiO2) nanoparticles have been utilized in the biomolecular recognition of antitumor or anticancer drug daunorubicin. Our observations of electrochemical studies indicate that because of the relative large surface area of the blends, the new nano TiO2-PLA polymer nanocomposites could remarkably increase the detection sensitivity as well as the binding affinity of daunorubicin to DNA, which is also demonstrated in our spectroscopic study. Besides, to further reveal the effect of the new TiO2-PLA polymer nanocomposites for the biorecognition, the relative influence of nano TiO2 or PLA themselves has also been explored in this study. The results illustrate that the drug molecules and/or DNA could be more readily self-assembled on the surface of the blends of the nano TiO2-PLA nanofibers so that the new nanocomposites could efficiently facilitate the relative biorecognition of daunorubicin.