Autism spectrum disorder (ASD) seriously affects children’s health. Recently, propionic acid (PA) has been reported to play a significant role in the formation of ASD. In this study, we investigated the community structure of PA-related bacteria in healthy and ASD children by isolation and culture, while a group of representative PA-related bacteria were identified and characterized based on colony morphological observation, physiological and biochemical tests, as well as living/dead cells staining analysis and 16S rRNA gene sequencing. The results showed that the number of PA-related bacteria in healthy children was >100-fold higher than that in ASD children, while compared to healthy children, greater diversity was found in PA-associated bacteria from ASD children. The sensitivity of the representative strains to PA was affected by bacterial species, PA concentration and incubation time. The decrease of pH value was found in PA-resistant Lactobacillus plantarum strain 6-1 but not in PA-sensitive Enterococcus lactis strain 4-1, while biofilm formation of both strains were unaffected by PA. Furthermore, PA inhibited the propagation of the two selected bacteria rather than killed them, while the greater inhibitory effect was observed on strain 4-1. Overall, the result is of great significance for revealing the role of PA-related bacteria in development of ASD.Copyright © 2020. Published by Elsevier Ltd.