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VETERINARY IMMUNOLOGY

Evaluation of Indirect Enzyme-Linked Immunosorbent Assays and IgG Avidity Assays Using a Protein A-Peroxidase Conjugate for Serological Distinction between Brucella abortus S19-Vaccinated and -Infected Cows

Ana C. A. M. Pajuaba, Deise A. O. Silva, José R. Mineo
Ana C. A. M. Pajuaba
Laboratory of Immunoparasitology, Institute of Biomedical Sciences, Federal University of Uberlândia, 38400-902 Uberlândia, MG, Brazil
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Deise A. O. Silva
Laboratory of Immunoparasitology, Institute of Biomedical Sciences, Federal University of Uberlândia, 38400-902 Uberlândia, MG, Brazil
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José R. Mineo
Laboratory of Immunoparasitology, Institute of Biomedical Sciences, Federal University of Uberlândia, 38400-902 Uberlândia, MG, Brazil
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  • For correspondence: jrmineo@ufu.br
DOI: 10.1128/CVI.00444-09
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ABSTRACT

This study aimed to evaluate the use of protein A-peroxidase (horseradish peroxidase [HRPO]) in indirect enzyme-linked immunosorbent assays (iELISAs) and IgG avidity assays for serological distinction between Brucella abortus S19-vaccinated and -infected cows. Four groups were analyzed: GI, 41 nonvaccinated seropositive cows; GII, 79 S19-vaccinated heifers analyzed at 3 months postvaccination; GIII, 105 S19-vaccinated cows analyzed after 24 months of age; and GIV, 278 nonvaccinated seronegative cows. IgG levels and avidity to B. abortus smooth lipopolysaccharide (S-LPS) were determined using anti-bovine IgG-HRPO or protein A-HRPO conjugates. Similar levels of IgG anti-S-LPS were found with GI using both conjugates. Lower IgG levels were detected with GII, GIII, and GIV using protein A-HRPO. Both conjugates showed high performance in discriminating GI from GIII, with high sensitivity (Se; 97.6%) and specificity (Sp; 97.1%). Protein A-HRPO was better in distinguishing GI from GIV (Se, 97.6%; Sp, 94.6%) and GI from GII (Se, 80.5%; Sp, 94.9%). Protein A-HRPO excluded a higher number of positive samples with GII and GIV. IgG avidity showed that protein A-HRPO, but not anti-IgG-HRPO, was able to distinguish nonvaccinated from vaccinated cattle, showing a higher avidity index (AI) with GI than with GII, with 78% of serum samples in GII showing an AI of <50%. Therefore, the iELISA using B. abortus S-LPS antigen and protein A-HRPO conjugate for preferential detection of the IgG2 subclass was shown to be suitable for serological distinction between S19-vaccinated and -infected cows. Also, antibodies generated after vaccination showed lower avidity, suggesting a role for the IgG2 subclass as an antibody of higher-affinity maturation after infection, constituting an additional tool for differentiating vaccinated from infected cattle.

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Evaluation of Indirect Enzyme-Linked Immunosorbent Assays and IgG Avidity Assays Using a Protein A-Peroxidase Conjugate for Serological Distinction between Brucella abortus S19-Vaccinated and -Infected Cows
Ana C. A. M. Pajuaba, Deise A. O. Silva, José R. Mineo
Clinical and Vaccine Immunology Mar 2010, 17 (4) 588-595; DOI: 10.1128/CVI.00444-09

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Evaluation of Indirect Enzyme-Linked Immunosorbent Assays and IgG Avidity Assays Using a Protein A-Peroxidase Conjugate for Serological Distinction between Brucella abortus S19-Vaccinated and -Infected Cows
Ana C. A. M. Pajuaba, Deise A. O. Silva, José R. Mineo
Clinical and Vaccine Immunology Mar 2010, 17 (4) 588-595; DOI: 10.1128/CVI.00444-09
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KEYWORDS

Antibodies, Bacterial
Antibody Affinity
Brucella Vaccine
Brucellosis
Cattle Diseases
Immunoglobulin G

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