Objective: To determine the diagnostic accuracy of Indian Scale for Assessment of Autism (ISAA) in children aged between 2-5 years. Design: Study of diagnostic accuracy Setting: Tertiary level hospital, (November 2015-November 2017). Participants: A consecutive sample of 500 children with suspected Autism (delay or regression of developmental milestones, delay or regression in speech, age-inappropriate understanding, behaviour, play and/or social interaction) was recruited. Procedure: Each child underwent an expert comprehensive assessment of Autism (reference tool) that included history, observation, examination, diagnostic criteria for Autism Spectrum Disorder (ASD) of the Diagnostic and Statistical Manual of Mental Disorders', 5th edition, Childhood Autism Rating Scale-2 (CARS2), developmental status and adaptive function. This was followed by the administration of ISAA (test tool) in Hindi language. Parameters of diagnostic accuracy and Receiver Operating Characteristic curves were computed. Main Outcome Measures: ASD based on (i) expert assessment, (ii) CARS-2, and (iii) ISAA. Results: In children aged 2-3 years, sensitivity of ISAA was 100% (95% CI 98.2% -100%), specificity 28.9% (95% CI 17.7% to 43.4%), positive likelihood ratio 1.4 and negative likelihood ratio 0. In 3-5 year olds, sensitivity was 99.6% (95% CI 97.6% to 99.6%), specificity 33.3% (95% CI 15.1% to 58.3%), positive likelihood ration 1.5 and negative likelihood ratio 0.01. The degrees of autism based on the existing cut off values were inaccurate. Conclusions: ISAA has sub-optimal performance in diagnosing and assessing severity in 2-5 year old children.