Campylobacteriosis -caused principally by Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli) - is among the main causes of bacterial gastroenteritis worldwide. This work was done to investigate the molecular characterization of zoonotic C. jejuni and C. coli isolated from fecal samples of beef cattle, retail beef meat and beef liver and stool of children with diarrhea. Fecal samples were collected from 50 apparently healthy cattle, 60 of retail beef meat and beef liver (30 of each) as well as 50 stool samples from pediatric diarrhea were subjected to standard isolation and phenotypic identification of Campylobacter isolates. The prevalence of Campylobacter isolate was 17(34%) in fecal sample of cattle, 5(16.66%) beef meat, 8(26.66 %) beef liver and 13 (26%) in pediatric diarrhea. Out of 43 identified isolates, 26(60.46%) C. jejuni isolates were higher than 14(32.55%) C. coli, two samples were mixed infection and one Campylobacter upsaliensis. A multiplex-PCR method was developed for the detection of C. jejuni and C. coli. Primers were the hippuricase gene (hipO) characteristic of C. jejuni, a sequence partly covering an aspartokinase gene (asp) characteristic of C. coli and a universal 16S rDNA gene sequence serving as an internal positive control. All Campylobacter isolates expressed identity with 16S rDNA (genus specific gene) at 1062 pb. Multiplex PCR demonstrated one false- positive and one false-negative hippurate activity test. PCR method was incapable to identify biochemically identified C. upsaliensis. Amplification of hipO gene of C. jejuni and asp- gene of C. coli isolated from cattle, beef and liver have shown identical fingerprints with human C. jejuni and C. coli at 344bp and 500bp respectively, indicating the public health importance of the isolates