Preparation of diagnostic polyclonal and monoclonal antibodies against outer envelope proteins of Serpulina pilosicoli

The purpose of this study was to prepare specific sera for use in the rapid detection and identification of the intestinal spirochaete Serpulina pilosicoli. In Western blot analysis, with pig antiserum which was raised against whole cells of S. pilosicoli and absorbed with outer envelope protein extracts from S. hyodysenteriae and S. innocens, a prominent protein with Mr of c. 72 kDa was consistently identified in outer envelope preparations of S. pilosicoli strains. Immunogold labelling demonstrated that this was located on the outer surface of intact S. pilosicoli cells. Two monoclonal antibodies (MAbs), designated C12 and M96, were raised against the protein. Although C12 reacted with a protein band of c. 72 kDa, this was also present in preparations from strains of other Serpulina spp. examined. MAb M96 reacted with an 80-kDa protein which was present only in preparations made from strains of S. pilosicoli. This was used in Western blot analysis and in an immunodot-blot assay with outer envelope extracts to specifically identify S. pilosicoli strains isolated from man, pigs, dogs and poultry. An indirect immunofluorescence test with MAb M96 also was used to detect and identify whole S. pilosicoli cells. Therefore, both the cross-absorbed antiserum and MAb M96 are potentially useful reagents for the detection and identification of S. pilosicoli.