The clinical and radiological outcomes of two-level anterior lumbar interbody fusion (ALIF) with percutaneous pedicle screw fixation (PSF) were evaluated in 24 consecutive patients who underwent two-level ALIF with percutaneous PSF for segmental instability and were followed up for more than 3 years. Clinical outcomes were assessed using a visual analogue scale (VAS) score and the Oswestry Disability Index (ODI). Sagittal alignment, bone union, and adjacent segment degeneration (ASD) were assessed using radiography and magnetic resonance imaging. The mean age of the patients at the time of operation was 56.3 years (range 39-70 years). Minor complications occurred in 2 patients in the perioperative period. At a mean follow-up duration of 39.4 months (range 36-42 months), VAS scores for back pain and leg pain, and ODI score decreased significantly (from 6.5, 6.8, and 46.9% to 3.0, 1.9, and 16.3%, respectively). Clinical success was achieved in 22 of the 24 patients. The mean segmental lordosis, whole lumbar lordosis, and sacral tilt significantly increased after surgery (from 25.1°, 39.2°, and 32.6° to 32.9°, 44.5°, and 36.6°, respectively). Solid fusion was achieved in 21 patients. ASD was found in 8 of the 24 patients. No patient underwent revision surgery due to nonunion or ASD. Two-level ALIF with percutaneous PSF yielded satisfactory clinical and radiological outcomes and could be a useful alternative to posterior fusion surgery.