Development of an ELISA to detect antibodies to a protective lipopolysaccharide fraction of Leptospira borgpetersenii serovar hardjo in cattle.
Monoclonal antibodies (Mabs) were produced against Leptospira borgpetersenii serovar hardjo-type Bovis antigens. A panel of 28 Mabs were characterised. Only the nine Mabs toward a lipopolysaccharide (LPS) fraction of 18, 24 kDa bands and a 26-28 kDa smear showed agglutinating, leptospiricidal and growth-inhibition activities, and passively protected hamsters against renal infection with hardjo. They also reacted strongly in the CH-ELISA, captured killed whole hardjo leptospires, gave good fluorescence in indirect FAT against smears of hardjo culture and exhibited no cross reactivity with strains in heterologous serogroups. On the basis of optimal activity in a range of tests, one IgG class Mab (designated 25) was selected for use in an antibody-capture ELISA system for the detection of bovine anti-hardjo antibodies. The system gave a wide separation of absorbance values between positive and negative sera at a 1:10 dilution. The antibodies detected by this assay are believed to be protective anti-LPS IgG.
Yan
K T
KT
Department of Veterinary Science, Faculty of Agriculture and Food Science, Queen's University of Belfast, Northern Ireland, UK.
Ellis
W A
WA
Mackie
D P
DP
Taylor
M J
MJ
McDowell
S W
SW
Montgomery
J M
JM
D000372:Agglutination Tests / Q000662:veterinary; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis / Q000097:blood* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D000942:Antigens, Bacterial / Q000097:blood; D015153:Blotting, Western / Q000662:veterinary; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000276:immunology*; D006224:Cricetinae; D004197:Disease Reservoirs / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D007668:Kidney / Q000382:microbiology; D007919:Leptospira / Q000254:growth & development / Q000276:immunology*; D007922:Leptospirosis / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D008070:Lipopolysaccharides / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D008070:Lipopolysaccharides