Abstract: Background and Objective: Streptococcus suis (S. suis) is one of the infectious agents that can cause a variety of symptoms of the disease in pigs. The Papua people live closely with pigs and the pigs raise extensively around their Honai (traditional home). The pigs are part of their way of life and part of their family. Pigs also play an important role in traditional ceremonies and also most of laboratories would probably misidentify an isolate of S. suis. The aim of this study was to identify of S. suis isolated from pigs in Papua. Methodology: This study was designed to isolate, identify and characterize of S. suis isolated from pig in various districts in Papua. The presence of 16s rRNA, glutamate dehydrogenase (gdh gene), muramidase released protein (mrp gene), capsular polysaccharide (cps gene) were performed by the Polymerase Chain Reaction (PCR) assay. Samples were swabbed from the tonsil of pigs and cultured an aerobically in sheep blood agarand identified by Gram staining, biochemical tests and the growth properties in liquid media as weel as Serum Soft Agar (SSA). Results: According to results, it have been identified that 54 isolates were positive of S. suis (100%), but only 30% of isolates fermented trehalose and 74% of salicin. Most of S. suis isolates showed α-haemolytic of 88.9% on sheep blood agar and only 11.1% formed β-haemolytic. About 40.7% isolates grew with clear supernatant and 59.3% isolates with turbid supernatant. In the serum soft agar, more then half isolates grew diffuse and less than 37% isolates with compact characteristic. Based on genotype characteristics, all isolates were positive for 16s rRNA and gdh gene, no more 28% isolates were positive for mrp and cps2j gene. Conclusion: This study indicated that all isolates had been correctly identified by the biochemical methods and that PCR was more definitive and revealed the inherent problem associated with the interpretation of biochemical results.