Immunobiochemical Techniques for Detection and Identification of Nanobacteria Isolated from Renal Stones
K. Abo-EL-Sooud , M.M. Hashem and A.Q. Gab-Allaha
Abstract: The present study is conducted to isolate on NB on solid media (Loeffler media) and to produce polyclonal antibodies for immunofluorescence assays. Moreover, to evaluate the re-isolation and molecular characterization of NB from kidney of experimentally infected mice. The culture of NB on solid media was visible macroscopically and microscopically as pin point at the bottom of the plat as stony like appearance with grayish to yellowish whit color after 6-8 weeks. While the result of DNA staining by Hoechst stain gave fluorescence appearance surrounding the NB which may be attributed to that the stain cannot penetrated the apatite layer on the surface of the NB. The histopathological examination showed the macroscopic enlargment and the inflamation of the kidney of infected mice while the microscopic examination revealed sever interstitial nephritis and thickening in parietal layer of Bowman’s capsule indicating the pathological action of NB. Finally the result of DNA extraction and Polymerase Chain Reaction (PCR) was negative. Results proved the positive identification of NB involved specific stainability with Hoechst 33258 and with indirect immunofluorescence staining (IFS) on both types of media. These tools will be promising in diagnosis criterion of NB.
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		How to Cite: K. Abo-EL-Sooud, M.M. Hashem and A.Q. Gab-Allaha , 2014. Immunobiochemical Techniques for Detection and Identification of Nanobacteria Isolated from Renal Stones . Insight Nanotechnology, 4: 1-7 DOI: 10.5567/INANO-IK.2014.1.7