Objective: This study was conducted to determine the clinical characteristics and differential diagnosis of individual cases of acute flaccid paralysis (AFP) reporting to tertiary care teaching hospital of Jaipur. Methods: This prospective observational study was conducted among patients attending outdoor, indoor, and neurology clinics with provisional diagnosis of AFP. The diagnosis was based on the available clinical data, vaccination status, and laboratory results (stool examination, arterial blood gas analysis, thyroid profile, serum electrolytes, electrophysiological studies, cerebrospinal fluid analysis, urinary pH, and imaging). Results: A total of 60 patients were evaluated over a study duration of 1 year. About 81.7% were male. Most common presenting complaint was lower-limb weakness (100%). None of the cases were identified as acute poliomyelitis or polio-compatible. Guillain–Barré syndrome (GBS) was the most common cause of non-poliovirus AFP (75%). Most common nerve conduction velocity findings in our study were bilateral motor axonal affection of common peroneal nerve (55%). Conclusion: The most common cause of AFP was GBS in all age groups. No case was diagnosed as poliomyelitis. All of the patients of GBS had progressive muscle weakness in a roughly symmetrical distribution, with areflexia of lower limbs.