Introduction: It has been suggested that viruses, especially herpesviruses, can play a role in the pathogenesis of marginal and apical periodontitis. This study aimed to detect herpesviruses types 1 to 8, namely herpes simplex virus (HSV-1/2), varicella zoster virus (VZV), Epstein-Barr virus (EBV), human cytomegalovirus (HCMV), human herpesvirus-6 (HHV-6), human herpesvirus-7 (HHV-7), and human herpesvirus-8 (HHV-8) as well as human papillomavirus (HPV) in acute apical abscesses. Methods: Twenty-four samples were taken by aspiration of the purulent exudate from acute apical abscesses. DNA extracted from clinical samples served as a template in single or nested polymerase chain reaction (PCR) assays for the detection of the target viruses. Results: Control PCR reactions with β-globin gene primers revealed that all samples but one had detectable human DNA. Of the 23 abscess samples positive for the β-globin gene, 14 (61%) were positive for at least one of the target human viruses. Thirteen (56.5%) cases had herpesvirus: HHV-8 occurred in 11 (48%), VZV and HHV-6B in two (9%), and HHV-7 and HSV-1/2 in one (4%). EBV and HCMV were not present in any of the examined samples. HPV was detected in three (13%) abscess samples. Viral coinfection was found in five cases, with one case harboring three of the targeted viruses. Conclusion: A large number of abscess samples were positive for at least one target virus. Unexpectedly, HHV-8 was for the first time detected and in a high prevalence. Papillomavirus and other herpesviruses were also found for the first time in endodontic abscesses. Although these findings suggest an association, the specific role of viruses in the pathogenesis of acute apical abscesses awaits further clarification.