Aims To evaluate the microbiota of endodontic infections in deciduous teeth by checkerboard DNA-DNA hybridization after uniform amplification of DNA in samples by multiple displacement amplification (MDA). Methodology Forty samples from the root canal system of deciduous teeth exhibiting pulp necrosis with or without radiographically detectable periradicular/interadicular bone resorption were collected and 32 were analyzed, with 3 individuals contributing 2 samples; these were MDA- amplified and analyzed by Checkerboard DNA-DNA hybridization for levels of 83 bacterial taxa. Two outcome measures were used: the percentage of teeth colonized by each species; and the mean proportion of each bacterial taxon present across all samples were computed. Results The mean amount of DNA in the samples prior to amplification was 5.2 (± 4.7) ng and 6.1 (± 2.3) μg after MDA. The mean number of species detected per sample was 19 (± 4) (range: 3–66) to the nearest whole number. The most prevalent taxa were Prevotella intermedia (96.9%), Neisseria mucosa (65.6%), Prevotella nigrescens (56.2%) and Tannerella forsythia (56.2%). Aggregatibacter (Haemophilus) aphrophilus and Helicobacter pylori were not detected. P. intermedia (10%), Prevotella tannerae (7%) and Prevotella nigrescens (4.3%) presented the highest mean proportions of the target species averaged across the positive samples. Conclusion Root canals of infected deciduous teeth had a diverse bacterial population. Prevotella sp were commonly found with P. intermedia, Prevotella tannerae and Prevotella nigrescens among the most prominent species detected.