A total of 250 samples including 100 samples from chicken meat, 75 samples human diarrhea, 75 samples from environment were collected from Aswan, Egypt. All samples were bacteriologically and biochemically examined for isolation, identification and differentiation of campylobacter spp., multiplex PCR detection of 23S rRNA, hipO, glyA gene for identification and differentiation of campylobacter spp. and detection of some pathogenic virulence genes include Iam, cdtB and cadF genes. Prevalence of campylobacter in chicken meat, human diarrhea and environmental samples by conventional methods were 32%, 14.7% and 13.3%, respectively. Prevalence of campylobacter in chicken meat, human diarrhea and environmental samples by mPCR were 6%, 5.33% and 4%, respectively. About 64.3%, 21.4% and 14.3 of examined samples were Campylobacter jejuni, Campylobacter coli and mixed culture, respectively. Most positive samples contain high prevalence of pathogenic virulence genes. Poultry meat and environment could be a dangerous source for pathogenic campylobacter for human. Most campylobacter isolates have a lot of pathogenic genes which increase the invasiveness and pathogenicity of Campylobacter.