Serodiagnosis and monitoring of contagious bovine pleuropneumonia (CBPP) with an indirect ELISA based on the specific lipoprotein LppQ of Mycoplasma mycoides subsp. mycoides SC.
An indirect ELISA, based on the specific and strongly antigenic recombinant peptide of the N'-terminal half of the lipoprotein LppQ from Mycoplasma mycoides subsp. mycoides small colony type (SC) was developed for the detection of antibodies to M. mycoides subsp. mycoides SC. It was evaluated for its suitability for serodiagnosis and monitoring of contagious bovine pleuropneumonia (CBPP). The recombinant peptide containing poly-histidine residue tails was expressed in Escherichia coli and subsequently purified by Ni(2+) chelate affinity chromatography to be used as antigen to coat microtiter ELISA plates. The specificity of the antigen was tested against rabbit hyperimmune sera directed against related Mycoplasmas of the M. mycoides cluster and with sera from cattle that were either free of CBPP, but suffered from other mycoplasmal infections such as M. bovis, or showed cross-reactions in the complement fixation test. The sensitivity of the ELISA was assessed with sera from artificially infected animals and with sera from cattle originating from areas where CBPP was endemic at the time of blood sampling. The study revealed that the ELISA was both specific and sensitive for CBPP positive bovine sera and was shown also to be robust to harsh climatic conditions.
Bruderer
Urs
U
Bommeli Diagnostics, Stationsstr. 12, CH-Liebefeld, Bern, Switzerland.
Regalla
Jose
J
Abdo
El-Mostafa
el-M
Huebschle
Otto J B
OJ
Frey
Joachim
J
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology; D002138:Calibration; D002417:Cattle; D002418:Cattle Diseases / Q000097:blood / Q000175:diagnosis*; D002980:Climate; D003168:Complement Fixation Tests / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D000939:Epitopes / Q000276:immunology; D007106:Immune Sera / Q000276:immunology; D008074:Lipoproteins / Q000276:immunology*; D009176:Mycoplasma mycoides / Q000276:immunology*; D011002:Pleuropneumonia, Contagious / Q000097:blood / Q000175:diagnosis*; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000662:veterinary
D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D000939:Epitopes; D007106:Immune Sera; D008074:Lipoproteins; D011994:Recombinant Proteins