Nanobacteria are the smallest described bacteria to date, with dimensions of 0.08 to 0.50 ìm. Furthermore, these organisms were found to produce a biofilm containing hydroxyl apatite or carbonate, preventing their effective staining. Theywere also isolated from commercial serumused in the cell culture. Nanobacteria have been detected thereafter in blood and blood products derived from horses, as well as blood from human blood donors. Some strains were isolated from kidney stones and bovine sera. To confirm the data, we searched for Nanobacteria fromaseptically removed urinary tract stoned.We used scanning electronmicroscopy for these stones. Our objectives are identification and determination of Nanobacteria by cell co culture and apatite formation by Nanobacteria in Loeffler medium and SF medium. Also production of polyclonal antibodies, immune-fluorescence assays. Moreover, identification using molecular characterization of Nanobacteria by PCR amplification of 16S rRNA gene of selected isolates and cloning for PCR amplified products. Microbiological characteristics:We have reviewed recent publications regarding the microbiological characteristic and pathogenicity of a novel infectious agent, the mineral-forming , sterile-filterable, slow-growing Gram-negative Nanobacteria, detected in bovine/human blood, kidney cyst fluid, urine and kidney stones. According to their 16S rDNA structure, nanobacteria belong to the alpha-2 Proteobacteria, subgroup, which includes the Brucella and Bartonella species. Their cell diameter is 0.2-0.5 microm (the smallest known cell-walled bacteria). Their most remarkable characteristic is the formation of carbonate apatite crystals of neutral pH and at physiologic phosphate and calcium concentrations. The extracellular mineralization forms a hard protective shelter for these hardy microorganisms, and enables them to survive conditions of physical stress that would be lethal to most other bacterial species.