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Dialister pneumosintes is an anaerobic Gram-negative rod that has been recently implicated as a candidate endodontic pathogen. In this study, samples taken from abscessed teeth and infected root canals associated with asymptomatic or symptomatic periradicular lesions were examined for the occurrence of bacterial associations involving D. pneumosintes. DNA was extracted from the samples, and the presence of D. pneumosintes and 16 other bacterial species was determined by means of species-specific nested polymerase chain reaction. Positive and negative associations involving D. pneumosintes were investigated by computing the odds ratio of D. pneumosintes being found in a sample from endodontic infection in co-infection with one of the other target species. The association between the pairs containing D. pneumosintes and the occurrence of pain also was evaluated. D. pneumosintes was always detected in mixed infections with at least two of the other target species. D. pneumosintes was positively associated with Treponema denticola, Porphyromonas endodontalis, Fusobacterium nucleatum, Peptostreptococcus micros, Campylobacter rectus, Prevotella intermedia, T. pectinovorum, and T. vincentii. Negative associations were observed with Bacteroides forsythus, P. gingivalis, and Actinomyces israelii. No pair containing D. pneumosintes was found to be significantly associated with symptomatic cases (p > 0.01). The findings of this study lend considerable support to the notion of D. pneumosintes being an important endodontic pathogen, usually in a mixed infection. Positive associations of this species with other highly prevalent species, such as T. denticola and P. endodontalis, suggest that bacterial synergism can occur and thereby play an important role in the pathogenesis of different forms of periradicular lesions.