Streptococcus and Enterococcus infections in chickens may result in significant negative effect on economy. In the current study, the prevalence of Streptococcus and Enterococcus species was planned in different broiler chickens farms in Beni-Suef Governorate. A total of 272 samples were collected from lesions (septicemic organs, enlarged organs, necrotic focci) of the affected organs including heart, lung, liver and kidney of diseased broiler chickens and freshly dead ones. Out of 272 samples a total of 49 isolates were recovered with incidence of 18% including 26Streptococcus spp. (53.1%) and 21 Enterococcus spp. (42.8%) meanwhile 2 isolates (4.1%) were unidentified. Streptococcus isolates were identified as S.gallinaceous (24.5%), S. dysgalactiae (16.3%) and S. durans (12.2%). Meanwhile all Enterococcus isolates were identified as E. faecalis. The in-vitro antibiotic sensitivity testing showed that all isolates were highly sensitive to amoxicillin (77.6%), sulfamethoxazole-trimethoprim (73.5%) and amoxicillin-clavulanic acid (65.3%). Meanwhile, all isolates were resistant to cephalexin, cefotaxime sodium, cefipime, cefotriaxone, tetracycline, kanamycin and apramycin while 87.8 and 63.2% of isolates showed resistance aganist gentamicin and enrofloxacin, respectively. Moreover, multidrug resistant were detected in all isolates. Polymerase chain reaction (PCR) was applied to identify 4 resistance-associated genes including (tetO, aac(6')aph(2''), blaZ and Pbp < /em>1A) as well as 6 virulence-associated genes including (cylE, brp < /em>A, hyl, cylA, asa1 and gelE). The results indicated that tetO, aac(6')aph(2''), blaZ, Pbp < /em>1A, cylE, brp < /em>A, cylA and asa1 genes were recovered from all the tested isolates (100%). Meanwhile, none of streptococcus isolates had hly gene also, gelE gene not detected in enterococcus isolates.