Abstract
Rabbit IgG raised against whole cells ofYersinia enterocolitica O:3, O:9 and against a group of pathogenicY. enterocolitica strains (serotypes O:3, O:5,27, O:8. and O:9) were prepared. The antibody limiting titers were within the range of 1:9.5 × 104−1:7.5 × 105. The immunoblotting analysis ofYersinia lipopolysacchides separated by SDS-PAGE showed that IgG against the single serotype O:3 interacted with high-molar-mass LPS of O:3 whereas other antibodies were bound to low-molar-mass LPS of serotypes O:3, O:5,27, O:9 and strainY. enterocolitica (CNCTC Y 2/68). IgG against the group of pathogenic serotypes also weakly interacted with low-molar-mass LPS of serotypes O:5, O:6,30, and O:10. The cross-reactivity of the antibodies withY. pseudotuberculosis Ia and/orY. rohdei b, d, e, f, i, which was observed by means of dot-blotting procedure using the whole bacterial cells as an antigen, was shown not to be caused by LPS of these bacteria. The prepared antibodies were used in the development of indirect competitive ELISA. At the optimum concentration of the immunoreactants the detection limits were within the range of 3–7 × 106 colony-forming units per mL.
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Abbreviations
- BSA:
-
bovine serum albumin
- CELISA:
-
competitive ELISA
- ELISA:
-
enzyme-linked immunosorbent assay
- IgG:
-
immunoglobulin G
- IPBS:
-
isotonic phosphate buffered saline, pH 7.0
- LPS:
-
lipopolysaccharide
- PAGE:
-
polyacrylamide gel electrophoresis
- PBS 7.4:
-
10 mmol/L phosphate-buffered saline, pH 7.4
- PBST:
-
PBS 7.4 containing 0.05 % Tween 20
- Px:
-
horseradish peroxidase
- SDS:
-
sodium dodecyl sulfate
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This study was supported by theGrant Agency of the Czech Republic (project GAČR 525/05/0311 —Rapid methods for detection of pathogenic Yersinia enterocoliticain foods).
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Hochel, I., Škvor, J. Characterization of rabbit antibodies for immunochemical detection ofYersinia enterocolitica . Folia Microbiol 52, 511–518 (2007). https://doi.org/10.1007/BF02932112
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DOI: https://doi.org/10.1007/BF02932112