An antigen capture ELISA test using monoclonal antibodies for the detection of Mycoplasma californicum in milk.
The use of monoclonal antibodies to detect Mycoplasma californicum was investigated in an antigen capture microtitre format. The finalized test was highly specific and no cross-reactions were detectable with any of the mastitis associated mycoplasma or bacterial antigens tested. Using a concentration step involving centrifugation, the sensitivity of the test could be improved from 10(5)-10(7) to 10(3)-10(5) colony forming units per ml with pure broth cultures, and from 10(7) to at least 10(6) colony forming units per ml in milk samples from two experimentally infected cows. The antigen detected was partially identified by immunoblotting, which demonstrated two polypeptides of 40 and 46 kD.
Veterinary Research Laboratories, Stormont, Belfast, Northern Ireland.
H J
HJ
Ball
D P
DP
Mackie
D
D
Finlay
J
J
McNair
D A
DA
Pollock
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000942:Antigens, Bacterial / Q000276:immunology / Q000302:isolation & purification*; D002417:Cattle; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D005260:Female; D015151:Immunoblotting / Q000662:veterinary; D008414:Mastitis, Bovine / Q000175:diagnosis; D051379:Mice; D008807:Mice, Inbred BALB C; D008892:Milk / Q000382:microbiology*; D009174:Mycoplasma / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial