Enzyme immunoassay for the diagnosis of brucellosis: chimeric Protein A-Protein G as a common enzyme labeled detection reagent for sera for different animal species.
A recombinant protein combining the immunoglobulin binding sites of Proteins A and G, conjugated with horseradish peroxidase was used as a universal detection reagent for the assessment of antibodies against Brucella spp. The reagent was applied in an indirect enzyme immunoassay for detection of antibodies to smooth lipopolysaccharide antigen in sera from Brucella spp. exposed and non-exposed cattle, sheep, goats and pigs and to antibodies to rough lipopolysaccharide in sheep, dogs and cattle. The results were similar to those obtained when murine monoclonal antibody-enzyme conjugates were used. An added advantage was that a universal cut-off for all tests using the proteins A and G detection reagent could be established, simplifying diagnostic interpretation of the data.
Nielsen
K
K
Animal Diseases Research Institute, Canadian Food Inspection Agency, 3851 Fallowfield Road, Nepean, Ont., Canada K2H 8P9. nielsenk@inspection.gc.ca
Smith
P
P
Yu
W
W
Nicoletti
P
P
Elzer
P
P
Vigliocco
A
A
Silva
P
P
Bermudez
R
R
Renteria
T
T
Moreno
F
F
Ruiz
A
A
Massengill
C
C
Muenks
Q
Q
Kenny
K
K
Tollersrud
T
T
Samartino
L
L
Conde
S
S
Draghi De Benitez
G
G
Gall
D
D
Perez
B
B
Rojas
X
X
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D002002:Brucella / Q000254:growth & development*; D002006:Brucellosis / Q000097:blood / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D002417:Cattle; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D005188:False Negative Reactions; D005189:False Positive Reactions; D006041:Goats; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D012680:Sensitivity and Specificity; D012756:Sheep; D013205:Staphylococcal Protein A / Q000737:chemistry*; D013552:Swine
D000907:Antibodies, Bacterial; C029152:G-substrate; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A