Abstract: Several advanced tests for detection of Ralstonia solanacearum phylotype II, sequevar I (race 3 biovar 2) in different habitats are nowadays developed. The immunofluorescent antibody staining techniques (IFAS), along with enzyme linked immunosorbent assay (ELISA) have been widely practiced. Using IFAS, however, under certain circumstances produced non-specific reactions. In the present study, a total of suspected 711 bacterial isolates recovered over four years period (2009-2012) of routine testing of potato crop in PBRP laboratories, Egypt, were subject to further determinations. All these isolates were tested by IFAS using internationally approved antiserum brand to detect R. solanacearum. These isolates were preliminary screened on Semi Selective Media of South Africa (SMSA) medium for presumptive detection of R. solanacearum. Colonies showing characteristics morphology of the pathogen in concern were scored for 206 isolates on SMSA medium. These characteristically screened isolates were physiologically and biochemically tested to confirm identity to R. solanacearum. A sum of 505 bacterial isolates devoiding the described characteristics of the pathogen were categorized into 7 groups according to their gram-staining reaction, cell morphology and cultural characteristics. Each group embraces a tentatively similar isolates according to motility, catalase and oxidase test(s) in principal. The results of Biolog identification revealed the following species of the cross-reacting bacteria that being affiliated to different families: Acinetobacter calcoaceticus, Aeromonas caviae, Moraxella osloensis, Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas corrugata, Pseudomonas mendocina, Pseudomonas tolaasii, Salmonella typhimurium, Serratia proteamaculans, Shewanella algae, Sphingobacterium multivorum and Bacillus maroccanus. The aforementioned identified bacteria were tentatively tested for the possible pathogenic potential to plant as shown by causing soft rot in potato tubers and inducing Hypersensitive Reaction (HR) in leaves of Burley tobacco plants.