Stray neutron distribution in a medical cyclotron vault room was evaluated by neutron activation analysis (NAA). Neutrons were generated in the production of radioactive nuclides, such as 18F, 11C, 13N and 15O, for diagnostic usage. Indium foil was adopted to evaluate the stray fast and thermal neutron intensity based on 115In(nf, n′)115mIn and 115In(nth, γ)116m1In reactions, respectively. The indium foils were weighed, sealed and placed at 62 points around the 6.7×8.2 m2 cyclotron room. Additionally, each indium foil was exposed for over 80 minutes during cyclotron operation and γ-peaks were analyzed using an HPGe detector to evaluate the number of stray fast (Φ f) or thermal (Φ th) neutrons. The minimum to maximum numbers of fast and thermal neutrons were (3.47±0.11)×103 to (1.06±0.21)×104 n·cm−2·s−1 and 9 to 965 n·cm−2·s−1, respectively. The minimum detectable limit for stray neutrons was included herein to demonstrate the reliability. Accordingly, 60 and two points, respectively, the confidence level associated with the reported intensities of fast and thermal neutrons reached 95%. The low qualified ratio in the evaluation of stray thermal neutrons might have been caused by either the high Compton scattering plateau or the low intensity of the gamma-ray peak in the relevant spectrum.