The present study was set to develop comprehensive system for assessing the safety of drinking water using PCR-reverse blot hybridization assay (REBA). The REBA developed in this study can detect waterborne pathogens such as Shigella spp., Salmonella spp., Citrobacter spp., Enterobacter spp., Klebsiella spp., Yersinia spp., Mycobacterium spp., Listeria spp. at the genus level, and Escherichia coli, Citrobacter freundii, Klebsiella pneumoniae, Pseudomonas aeruginosa, Yersinia enterocolitica, Y. pseudotuberculosis, Mycobacterium avium complex, M. marinum, Enterococcus faecalis, and Staphylococcus aureus at the species level, and E. coli O157:H7 at the strain level.