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

Nielsen, Klaus; Smith, Phillip D.; Wang, Yu; Nicoletti, P; Elzer, Philip H.; Vigliocco, Ana M.; Silva, P.; Bermudez, R.; Renteria, T.; Moreno, Francisco J. Magraner; Ruiz, Andrés M.; Massengill, C; Muenks, Q.; Kenny, Kevin; Tollersrud, Tore; Samartino, Luis; Conde, Simara Rufatto; Benitez, G Draghi de; Gall, D.; Pérez, Bárbaro Agustín Armas; Rojas, X

doi:10.1016/j.vetmic.2004.02.014

Cited by 34 publications

(19 citation statements)

References 21 publications

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“…The lack of availability of polyclonal or monoclonal antibodies raised to detect specifically the immunoglobulin isotypes of wildlife species was overcome by using protein G as a conjugate. This reagent has been reported suitable in wildlife for detecting antibodies to Brucella [52,53] and other pathogens [54,55]. Due to the absence of gold standard sera from culture positive and brucellosis free wild animals, we validated our iELISA using gold standard sera from the closest phylogenetically related domestic species.…”

Section: Discussionmentioning

confidence: 99%

Spatial distribution and risk factors of Brucellosis in Iberian wild ungulates

et al. 2010

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BackgroundThe role of wildlife as a brucellosis reservoir for humans and domestic livestock remains to be properly established. The aim of this work was to determine the aetiology, apparent prevalence, spatial distribution and risk factors for brucellosis transmission in several Iberian wild ungulates.MethodsA multi-species indirect immunosorbent assay (iELISA) using Brucella S-LPS antigen was developed. In several regions having brucellosis in livestock, individual serum samples were taken between 1999 and 2009 from 2,579 wild bovids, 6,448 wild cervids and4,454 Eurasian wild boar (Sus scrofa), and tested to assess brucellosis apparent prevalence. Strains isolated from wild boar were characterized to identify the presence of markers shared with the strains isolated from domestic pigs.ResultsMean apparent prevalence below 0.5% was identified in chamois (Rupicapra pyrenaica), Iberian wild goat (Capra pyrenaica), and red deer (Cervus elaphus). Roe deer (Capreolus capreolus), fallow deer (Dama dama), mouflon (Ovis aries) and Barbary sheep (Ammotragus lervia) tested were seronegative. Only one red deer and one Iberian wild goat resulted positive in culture, isolating B. abortus biovar 1 and B. melitensis biovar 1, respectively. Apparent prevalence in wild boar ranged from 25% to 46% in the different regions studied, with the highest figures detected in South-Central Spain. The probability of wild boar being positive in the iELISA was also affected by age, age-by-sex interaction, sampling month, and the density of outdoor domestic pigs. A total of 104 bacterial isolates were obtained from wild boar, being all identified as B. suis biovar 2. DNA polymorphisms were similar to those found in domestic pigs.ConclusionsIn conclusion, brucellosis in wild boar is widespread in the Iberian Peninsula, thus representing an important threat for domestic pigs. By contrast, wild ruminants were not identified as a significant brucellosis reservoir for livestock.

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Section: Discussionmentioning

confidence: 99%

Spatial distribution and risk factors of Brucellosis in Iberian wild ungulates

et al. 2010

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“…Thus, there is a need for a suitable substitute with ample specificity and sensitivity for detection of fish Ig. Previous studies in veterinary medicine have reported the use of enzyme-labelled protein A in serological studies of mammalian diseases such as brucellosis [23], Hantavirus disease [24] and Lyme disease [25]. Here, the usefulness of horseradish peroxidase conjugated protein A (protein A-HRP) in comparison to a mouse polyclonal antibody raised against purified rockfish Ig was evaluated for detection of L. garvieae in black rockfish, using 2-DE immunoblot and ELISA.…”

Section: Introductionmentioning

confidence: 99%

Efficacy of protein A-HRP in an immunological study of black rockfish (Sebastes schlegeli Higendorf) humoral immune responses

Kang

Shin

Palaksha

et al. 2006

Fish & Shellfish Immunology

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“…There is not a serological test appropriated for all the epidemiological situations; therefore, all the factors influencing in the relevance of the analytical method and the test results for a particular diagnostic interpretation or application must be taken into account. In the case of the rapid lateral flow immunochromatographic tests, even when they are simple and fast running systems, reliable and accurate results are reported (Smits et al, 2003;Genç et al, 2012;Díaz et al, 2015), which, together with the possibility of carrying them to the fields where the herds are (as field tests), make them an alternative to the conventional serological diagnosis of brucellosis (Smits et al, 2003;Nielsen et al, 2004). In the development of the LFIA-PG system, the appropriate concentration of the protein G conjugate was obtained for printing on the pads, and the migration buffer and drying time at 37 °C were effective.…”

Section: Resultsmentioning

confidence: 99%

“…Protein A reacts more specifically with the IgG 2 subclass, whereas protein G reacts with both subclasses of bovine IgG (IgG 1 and IgG 2), which allows detecting antibodies at any time of infection (Díaz et al, 2015;Shome et al, 2015). In comparative immunological studies on affinity and specificity of proteins A and G, protein G showed a greater affinity for G-type bovine immunoglobulins (Nielsen et al, 2004;Pajuaba et al, 2010;Genç et al, 2012). The present study showed the greater affinity of protein G for bovine immunoglobulins compared with protein A, observed in the results of sensitivity and specificity obtained by the LFIA-PG system.…”

Section: Resultsmentioning

confidence: 99%

Evaluation of two rapid immunochromatographic tests for diagnosis of brucellosis infection in cattle

Quintero¹,

Herrera

Alfonso³

et al. 2018

Open Vet J.

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Brucellosis is a serious infectious disease that causes significant economic losses in the livestock industry. Its early diagnosis allows an adequate disease control in cattle. DAVIH Laboratories designed a lateral flow immunochromatographic assay using protein A-colloidal gold as a detector reagent (LFIA-PA). The objective of this work was to compare the performance of this assay using protein G-colloidal gold (LFIA-PG) with its performance using protein A-colloidal gold as the detector reagent. The assays were carried out with 20 μL of serum and 130 μL of running buffer. Interpretation of bands was by visual inspection with the naked eye at 15- 20 minutes after sample application. The tests were evaluated with 449 samples of bovine serum (111 positive and 338 negative). The diagnostic sensitivity and specificity, the positive and negative predictive values, and the efficacy of both assays were calculated, and their concordance was estimated by calculating the kappa (k) index. The estimated values of the parameters for LFIA-PG and LFPIA-PA were 100% and 95.2% of diagnostic sensitivity, 96.2% and 97.3% of diagnostic specificity, 89.5% and 92.3% for the positive predictive value, 100% and 98.5% for the negative predictive value, and 97.1% and 96.89% of efficacy, respectively. The concordance between both tests was very good (k = 0.95). It was shown the possibilities of developing a system with LFIA-PG capable of detecting antibodies against Brucella spp. The performance of the test makes possible its use as a screening method in the diagnosis of brucellosis.

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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

Cited by 34 publications

References 21 publications

Spatial distribution and risk factors of Brucellosis in Iberian wild ungulates

Spatial distribution and risk factors of Brucellosis in Iberian wild ungulates

Efficacy of protein A-HRP in an immunological study of black rockfish (Sebastes schlegeli Higendorf) humoral immune responses

Evaluation of two rapid immunochromatographic tests for diagnosis of brucellosis infection in cattle

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