Expression of the SmpA outer membrane lipoprotein of Serpulina hyodysenteriae strain P18A in vivo

An ELISA has been developed using a monoclonal antibody (F325 AC4) to the SmpA surface lipoprotein of Serpulina hyodysenteriae strain P18A when grown in vitro. The lower level of detection of the ELISA was approxiamately 5 × 10 6 spirochaetes/ml when spirochaetes were either resuspended in phosphate...

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Bibliographic Details
Published in:	Veterinary microbiology 1995-04, Vol.44 (1), p.25-35
Main Authors:	Sellwood, R., Walton, F., Thomas, W., Burrows, M.R., Chesham, J.
Format:	Article
Language:	eng
Subjects:	Animals Antibodies, Monoclonal Bacterial Outer Membrane Proteins - analysis Bacterial Outer Membrane Proteins - biosynthesis Bacteriology Biological and medical sciences Blotting, Western - methods Brachyspira hyodysenteriae - growth & development Brachyspira hyodysenteriae - metabolism Brachyspira hyodysenteriae - pathogenicity CERDO DIAGNOSIS DIAGNOSTIC DIAGNOSTICO DISENTERIA PORCINA DYSENTERIE DU PORC Dysentery - diagnosis Dysentery - microbiology Dysentery - veterinary ELISA Enzyme-Linked Immunosorbent Assay - methods Feces - microbiology Fundamental and applied biological sciences. Psychology Gastrointestinal Contents - microbiology LIPOPROTEINAS LIPOPROTEINE LIPOPROTEINS Lipoproteins - analysis Lipoproteins - biosynthesis Microbiology Morphology, structure, chemical composition Outer membrane protein PORCIN Sensitivity and Specificity Serpulina hyodysenteriae SPIROCHAETALES Spirochaetales Infections - diagnosis Spirochaetales Infections - microbiology Spirochaetales Infections - veterinary SWINE Swine Diseases SWINE DYSENTERY TEST ELISA
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Summary:	An ELISA has been developed using a monoclonal antibody (F325 AC4) to the SmpA surface lipoprotein of Serpulina hyodysenteriae strain P18A when grown in vitro. The lower level of detection of the ELISA was approxiamately 5 × 10 6 spirochaetes/ml when spirochaetes were either resuspended in phosphate buffered saline or in pig faeces. When pigs were challenged with S. hyodysenteriae strain P18A the lipoprotein was detected in the faeces of pigs by ELISA when the numbers of spirochaetes excreted was greater than 10 6 per g of faeces. After onset of clinical signs in the pig, expression of SmpA was not detected by ELISA or by Western blotting using either monoclonal antibody F325 AC4 or polyclonal antiserum B50 against the SmpA antigen. However, when the in vivo grown spirochaetes were subsequently cultured in vitro expression of SmpA was detected by Western blotting. In the mouse model of swine dysentery S. hyodysenteriae spirochaetes obtained from mice with gross lesions also did not express SmpA. It was concluded that the apparent lack of expression mayhave been the result of environmental regulation of gene expression or antigenic variation and was not due to denaturation of the antigen in vivo.
ISSN:	0378-1135 1873-2542