Free-standing TiO2-SiO2/polyaniline (TS/PANI) composite nanofibers were prepared by electrospinning, in situ polymerization and calcination method. The effect of tetra-n-butyl titanate (TBT) in the electrospinning solution on the morphology and the ammonia sensing properties of TS/PANI composite nanofibers were investigated. The obtained nanofibers were characterized by scanning electron microscope, Fourier transform infrared spectroscopy, X-ray diffraction, transmission electron microscopy, thermo-gravimetric analysis and gas sensor test system. It is proved that too much TBT in the solution would make the fibrous morphology and ammonia sensing properties worse. Gas sensing tests showed that the TS/PANI composite nanofibers ammonia sensor can work at room temperature and possess ideal response values, selectivity and repeatability. With the increase in TiO2 content in the TS nanofibers, the ammonia sensing properties were improved because of the increase in P-N heterojunctions formed between TiO2 and PANI in the sensors.