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The aim of this in vitro study was to assess the antimicrobial efficacy of materials (gutta percha [GP], polytetrafluoroethylene [PTFE] tape, and vinyl polysiloxane [VP]) used for sealing the implant abutment screw hole (IASH).Sixty implants with a Morse cone connection were used. The implants were randomly divided into 3 groups (20 implants/group). In groups 1, 2, and 3, the IASH was sealed with GP, PTFE, and VP, respectively. Unstimulated whole saliva (UWS) was used as a contaminant medium for the incubation of the specimens. After 7 days of incubation, the specimens were opened under aseptic conditions, and the contents of the inner parts of the IASH were collected using sterile microbrushes. The occurrence of microbial leakage through the screw access channel interface was assessed with the checkerboard DNA-DNA hybridization method. P < 0.05 was considered statistically significant.Among samples in groups 1, 2, and 3, the UWS collected before incubation showed no statistically significant differences in relation to the prevalence of species investigated. Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Candida albicans were isolated from group 1 than groups 2 and 3.The choice of material for sealing the IASH is entirely dependent on the operators' choice and experience.