Development of an immunomagnetic antigen capture system for detecting leptospires in bovine urine.
A magnetic bead antigen capture system which combined the use of two evolving techniques - immunomagnetic separation (IMS) and time-resolved fluoroimmunoassay (TR-FIA) - was developed to detect Leptospira borgpetersenii serovar hardjo in bovine urine. The assay utilised monoclonal antibody coated magnetic beads to capture leptospiral antigen which was in turn detected using another monoclonal antibody (Indicator) labelled with biotin. Signal was generated by the binding of europium labelled streptavidin to indicator antibody. The sensitivity of the assay was improved from 10(3) to 10(2) leptospires per ml by using an ethanol precipitation procedure to treat each sample. The assay detected only 31 of 56 (55 per cent) urine specimens culture-positive for hardjo, but seven of 24 urine samples culture-negative for hardjo were identified as positive by the assay. These seven samples were from animals which were culture positive on at least one other occasion. These results suggest that this system should be further investigated as a complementary test to culture for the identification of hardjo carrier animals.
Yan
K T
KT
Department of Veterinary Science, Queen's University of Belfast, Northern Ireland.
Ellis
W A
WA
Montgomery
J M
JM
Taylor
M J
MJ
Mackie
D P
DP
McDowell
S W
SW
D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000652:urine; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D018189:Immunomagnetic Separation; D007919:Leptospira / Q000302:isolation & purification*; D007922:Leptospirosis / Q000175:diagnosis / Q000652:urine / Q000662:veterinary*; D008297:Male; D014556:Urine / Q000382:microbiology*
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial