Background/Aim: Understanding the composition of bacteria in infected root canals is important for ameliorating the treatment strategies that lead to the elimination of pathogens and infection control, but also prevent reinfection. Aim of this study was to investigate microbial composition of primary infected root canals with apical periodontitis of young permanent teeth, originating form school children in Serbia, and its association with clinical symptoms. Material and Methods: To determine the bacterial composition of infected root canals in children, 35 endodontic samples were obtained. The identification of cultured bacteria was performed by MALDI-TOF MS analysis. The presence or absence of clinical symptoms were recorded. Results: Facultative anaerobes were 2,2 times more frequent than obligate anaerobes. The most common facultative anaerobes belonged to following genera, Streptococcus (58 isolates), Actinomyces (10) and Enterococcus (8), while predominant obligate anaerobes, belonged to genera Veillonella (15), Prevotella (9) and Fusobacterium (8). The most common clinical isolates recovered from infected root canals with symptomatic apical periodontitis were Veillonella parvula (10) and Fusobacterium nucleatum (7), while from the asymptomatic ones, they were Streptococcus mitis/Streptococcus oralis (5). Prevalence of Parvimonas micra, Prevotella buccae and Streptococcus constellatus within the root canals might be associated to clinical symptoms. Conclusions: Species of genera Streptococcus and Veillonella were the most common isolates from primary infected root canals with apical periodontitis in Serbian school children. Facultative anaerobes were predominant over obligate anaerobes. The prevalence of obligate anaerobes was much higher in symptomatic compared to asymptomatic root canal infections. No specific bacterial strain might be associated to a single examined clinical symptom (pain, tenderness to percussion or swelling), but majority of the strains are associated to all of the examined three symptoms.