Development of a capture ELISA for the detection of antibodies to enteropathogenic Escherichia coli (EPEC) in rabbit flocks using intimin-specific monoclonal antibodies.
A capture enzyme-linked immunosorbent assay (cELISA) was developed using intimin-specific monoclonal antibodies to detect specific antibody in rabbits that have been in contact with enteropathogenic Escherichia coli (EPEC). Sera from 121 EPEC-negative, minimum-disease-level (MDL) rabbits were used for negative controls, and sera from 25 MDL rabbits, experimentally infected with EPEC of bio-/serotype 3-/O15, for positive controls. These were used to determine a cut-off value for a positive cELISA result. The value selected gave the test a sensitivity of 80.0% and a specificity of 98.4% on an individual level. At this value, a flock level sensitivity and specificity of 79.2 and 85.2%, respectively were calculated for a flock with a prevalence of seven per cent, if 40 animals were tested, and a minimum of two reactors were obtained. The test characteristics improve with increasing prevalence. To evaluate the diagnostic potential of the cELISA, sera from 40 to 50 slaughter rabbits per flock from 25 rabbit flocks with bacteriologically determined EPEC status were tested. The results demonstrated that this test can be a useful tool to determine the EPEC status of a rabbitry, provided that it is used at regular intervals.
Vandekerchove
D G F
DG
Department of Small Stock Pathology, Veterinary and Agrochemical Research Centre, Groeselenberg 99, 1180, Brussels, Belgium. dovan@var.fgov.be
Kerr
P G
PG
Callebaut
A P
AP
Ball
H J
HJ
Stakenborg
T
T
Mariën
J
J
Peeters
J E
JE
D018829:Adhesins, Bacterial / Q000276:immunology*; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins / Q000276:immunology*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D004926:Escherichia coli / Q000254:growth & development / Q000276:immunology / Q000302:isolation & purification*; D004927:Escherichia coli Infections / Q000175:diagnosis / Q000453:epidemiology / Q000382:microbiology / Q000662:veterinary*; D029968:Escherichia coli Proteins; D013058:Mass Spectrometry / Q000662:veterinary; D051379:Mice; D015995:Prevalence; D011817:Rabbits / Q000382:microbiology*; D012680:Sensitivity and Specificity
D018829:Adhesins, Bacterial; D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; C073819:eaeA protein, E coli