Objective To investigate the distribution and drug resistance of pathogenic bacteria isolated and cultured from valve vegetations in infective endocarditis, and to provide evidence for diagnosis and treatment of infective endocarditis in this area. Methods A total of 185 patients with infective endocarditis and cardiac surgery admitted to our hospital from March 2016 to December 2022 were selected to collect clinical data. Pathogens were isolated from the culture of valvular vegetations, and drug susceptibility tests were performed on common pathogens. Results The positive rate of valvular vegetation culture was 53.5% (99/185), and the pathogenic bacteria isolated from valvular vegetation culture included 70 strains of Streptococcus spp. (70.7%, 70/99) and 17 strains of Staphylococcus spp. (17.2%, 17/99), 5 strains of nutritionally variant Streptococci (5.0%, 5/99), 2 strains of Enterococcus spp. (2.0%, 2/99), 2 strains of HACEK organisms (2.0%, 2/99), 1 strain of Brucella ovis (1.0%, 1/99), 1 strain of Gemella haemolysans (1.0%, 1/99) and 1 strain of Pseudomonas (1.0%, 1/99). Streptococci spp. was completely sensitive to penicillin, cefotaxime, cefepime, vancomycin, meropenem, daptomycin and linezolid, while was partially resistant to tetracycline (47.1%, 33/70), erythromycin (38.6% , 27/70), clindamycin (27.1%, 19/70), levofloxacin (7.1%, 5/70) and chloramphenicol (1.4%, 1/70). Staphylococci spp. was completely sensitive to vancomycin, teicolanin, dattomycin, minocycline, chloramphenicol and linezolid, with resistance rates of 100%, 76.5%, 41.2% and 35.3% to penicillin, methicillin, erythromycin and clindamycin, respectively, and 10% to 20% to other commonly used drugs. Conclusions The main pathogens of infective endocarditis were Streptococcus spp. and Staphylococcus. spp. Positive valvular vegetation culture is valuable in supplementing and confirming blood culture. Penicillin remains the recommended treatment for streptococcal IE, and vancomycin can be a viable option for severe infections of Streptococcus spp. and Staphylococcus spp.