The ban on the use of antibiotics as growth promoters for poultry production in many countries has led to increasing interest to use probiotics as a promising alternative. Thus, the aim of this study is to isolate, screen and identify probiotics from poultry intestines, and to select probiotic candidates for subsequent in vivo experiment. A total of 60 lactic acid bacterial isolates were recovered from 21 to 42 days-old chickens onto De Man, Rogosa and Sharpe agar plates, then screened for their probiotic features using eleven in vitro assays. Among 60 LAB isolates, only three (SI5, IL22, and CE55) showed excellent probiotic features such as cell surface hydrophobicity, pH tolerance, survival in bile salts, cholesterol assimilation, NaCl tolerance, auto-aggregation, co-aggregation, hemolytic activity, antimicrobial activity, and lysozyme resistance as well as antibiotic susceptibility tests. The FASTA homology showed that the16S rRNA gene sequences of the selected isolates had 95.45, 99.88 and 100% nucleotide similarity with that of Enterococcus faecium NBRC 100486, E. faecium NBRC 100486 and E. faecium DSM 20477strains, respectively. Our results also suggest that new three strains have potential for future application as probiotic in health promoting foods and have the potential to enhance the immunity of infant against invading pathogenic microbes.