Skip to main content
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My Cart

Main menu

  • Home
  • Articles
    • Archive
  • About the Journal
    • About CVI
    • For Librarians
    • For Advertisers
    • FAQ
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My Cart

Search

  • Advanced search
Clinical and Vaccine Immunology
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Archive
  • About the Journal
    • About CVI
    • For Librarians
    • For Advertisers
    • FAQ
Article

Enzyme Immunoassays for Serological Diagnosis of Bovine Brucellosis: A Trial in Latin America

D. Gall, A. Colling, O. Marino, E. Moreno, K. Nielsen, B. Perez, L. Samartino
D. Gall
Canadian Food Inspection Agency, Animal Diseases Research Institute, Nepean, Ontario, Canada K2H 8P9 1 ;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
A. Colling
International Atomic Energy Agency, A1400, Vienna, Austria 2 ;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
O. Marino
Instituto Colombiano Agropecuaria, ICA-CEISA, Santafe de Bogota DC, Colombia 3 ;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
E. Moreno
Tropical Diseases Research Program, School of Veterinary Medicine, National University, Heredia, Costa Rica 4 ;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
K. Nielsen
Canadian Food Inspection Agency, Animal Diseases Research Institute, Nepean, Ontario, Canada K2H 8P9 1 ;
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
B. Perez
Servicios Agricolas y Ganaderos, Laboratorio Regional Osorno, Mackenna 674, Osorno, Chile 5 ; and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
L. Samartino
Instituto de Patobiologia-DPTO Bacteriologia, INTA-CICV, CC 77, 1708 Moron, Buenos Aires, Argentina 6
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Tables
  • Fig. 1.
    • Open in new tab
    • Download powerpoint
    Fig. 1.

    ROC curves representing each ELISA for data from Argentina. The cutoff value for each assay is indicated in the upper right corner of each panel. The AUC is indicative of how well the test performed. A value of 1.0 is perfect, and a value below the diagonal line represents reactivity due to chance. (a) IELISA-ADRI, AUC = 0.983. (b) IELISA-IAEA, AUC = 0.983. (c) CELISA-sLPS, AUC = 0.995. (d) CELISA-OC, AUC = 0.991.

  • Fig. 2.
    • Open in new tab
    • Download powerpoint
    Fig. 2.

    ROC curves representing each ELISA for data from Chile. (a) IELISA-ADRI, AUC = 1.000. (b) IELISA-IAEA, AUC = 0.996. (c) CELISA-sLPS, AUC = 1.000. (d) CELISA-OC, AUC = 1.000. See the Fig. 1 legend for additional explanation of the data.

  • Fig. 3.
    • Open in new tab
    • Download powerpoint
    Fig. 3.

    ROC curves representing each ELISA for data from Colombia. (a) IELISA-ADRI, AUC = 1.000. (b) IELISA-IAEA, AUC = 1.000. (c) CELISA-sLPS, AUC = 0.994. (d) CELISA-OC, AUC = 0.999. See the Fig. 1 legend for additional explanation of the data.

  • Fig. 4.
    • Open in new tab
    • Download powerpoint
    Fig. 4.

    ROC curves representing each ELISA for data from Costa Rica. (a) IELISA-ADRI, AUC = 0.992. (b) IELISA-IAEA, AUC = 0.991. (c) CELISA-sLPS, AUC = 0.974. (d) CELISA-OC, AUC = 0.969. See the Fig. 1 legend for additional explanation of the data.

  • Fig. 5.
    • Open in new tab
    • Download powerpoint
    Fig. 5.

    ROC curves representing each combined ELISA for data from Argentina, Chile, Colombia, and Costa Rica. (a) IELISA-ADRI, AUC = 0.985. (b) IELISA-IAEA, AUC = 0.989. (c) CELISA-sLPS, AUC = 0.995. (d) CELISA-OC, AUC = 0.995. See the Fig. 1legend for additional explanation of the data.

  • Fig. 6.
    • Open in new tab
    • Download powerpoint
    Fig. 6.

    Frequency distribution of ELISA data from Argentina. Open bars, serologically negative samples; closed bars, serologically positive samples. The numbers in each class limit are indicated on top of the bars. (a) IELISA-ADRI. (b) IELISA-IAEA. (c) CELISA-OC. (d) CELISA-sLPS.

  • Fig. 7.
    • Open in new tab
    • Download powerpoint
    Fig. 7.

    Frequency distribution of ELISA data from Chile. (a) IELISA-ADRI. (b) IELISA-IAEA. (c) CELISA-OC. (d) CELISA-sLPS. See the Fig. 6 legend for additional explanation of the data.

  • Fig. 8.
    • Open in new tab
    • Download powerpoint
    Fig. 8.

    Frequency distribution of ELISA data from Colombia. (a) IELISA-ADRI. (b) IELISA-IAEA. (c) CELISA-OC. (d) CELISA-sLPS. See the Fig. 6 legend for additional explanation of the data.

  • Fig. 9.
    • Open in new tab
    • Download powerpoint
    Fig. 9.

    Frequency distribution of ELISA data from Costa Rica. (a) IELISA-ADRI. (b) IELISA-IAEA. (c) CELISA-OC. (d) CELISA-sLPS. See the Fig. 6 legend for additional explanation of the data.

  • Fig. 10.
    • Open in new tab
    • Download powerpoint
    Fig. 10.

    Frequency distribution of combined ELISA data from Argentina, Chile, Colombia, and Costa Rica. (a) IELISA-ADRI. (b) IELISA-IAEA. (c) CELISA-OC. (d) CELISA-sLPS. See the Fig. 6 legend for additional explanation of the data.

Tables

  • Figures
  • Table 1.

    Comparison of procedures used for the two IELISAs and two CELISAsa

    ParameterAssay
    IAEA-IELISAADRI-IELISACELISA-OCCELISA-sLPS
    MicroplateNunc PolysorbNunc 69620Nunc 69620Nunc 69620
    AntigensLPSsLPSO chainsLPS
     Concn (μg/ml)1.01.02.01.0
     Buffer0.05 M CO3 0.05 M CO3 0.05 M CO3 0.05 M CO3
     Incubation temp (°C)4204 (frozen)20
     Incubation time (h)≥18≥18≥18≥18
    Wash buffer0.002 M PO4, 0.15 M NaCl, 0.05% Tween 20, pH 7.40.01 M PO4, 0.15 M NaCl, 0.05% Tween 20, pH 7.20.01 M PO4, 0.15 M NaCl, 0.05% Tween 20, pH 7.20.01 M PO4, 0.15 M NaCl, 0.05% Tween 20, pH 7.2
    No. of wash cycles3444
    Serum diluent0.01 M PBS, 0.05% Tween 20, pH 7.4Wash buffer plus EDTA-EGTA, pH 6.3Same as wash bufferWash buffer plus EDTA-EGTA, pH 6.3
    Serum (control/test)
     Serum dilution1:2001:501:501:20
     Incubation temp (°C)37202020
     Incubation time (min)603012030
     AgitationYesNo3 min3 min
    Competing antibodyNANAYsT9-HRPM84
    Detecting antibodyMAb to bovine IgG1-HRPMAb to bovine IgG1-HRPSame as competingGaMIgG-HRP (diluted in wash buffer)
     Incubation time (min)6030NA30
     Incubation temp (°C)3720NA20
     AgitationYesNoNANo
    Substrate/chromogen
     Incubation time (min)10101010
     Incubation temp (°C)37202020
     AgitationYesYesYesYes
     Wavelength (nm)405414414414
    • ↵a Abbreviations: PBS, phosphate-buffered saline; NA, not applicable; HRP, horseradish peroxidase; MAb, monoclonal antibody; IgG1, immunoglobulin G1; GaM, goat anti-mouse IgG-HRP.

  • Table 2.

    Number of samples tested in each country in each group (not exposed, serologically positive, or vaccinated withB. abortus)

    StatusaNo. of observations per country and combined
    ArgentinaChileColombiaCosta RicaCombined
    Negative2159725548722,613
    Positive7096922661901,857
    VaccinateNAb 9541,1101,0023,066
    • ↵a All samples were defined relative to their RBT and CFT results.

    • ↵b NA, data not available. There was an insufficient number of vaccinates to be a separate category, and so the data was combined with negatives.

  • Table 3.

    Comparison of percent positive reactions in each country with actual sensitivity determined in the Canadian study

    Country% for assay
    IELISA-ADRIIELISA-IAEACELISA-OCCELISA-sLPS
    Argentina92.6697.8896.9097.74
    Canada100NAb NA100
    Chile98.9997.1198.84100
    Colombia10010099.2598.12
    Costa Rica10098.4292.1093.16
    Combineda 96.7796.2897.0497.47
    • ↵a Combined data for all the countries except Canada (included for comparison).

    • ↵b NA, these tests were not part of the original Canadian study.

  • Table 4.

    Comparison of percent negative reactions in each country with specificity determined in the Canadian study

    Country% for assay
    IELISA-ADRIIELISA-IAEACELISA-OCCELISA-sLPS
    Argentina96.2898.1498.1498.14
    Canada99.40NAb NA99.90
    Chile99.2899.5999.6999.59
    Colombia99.8299.8299.8297.11
    Costa Rica95.7694.9593.3595.76
    Combineda 93.5797.0198.0598.32
    • ↵a Combined data for all the countries except Canada.

    • ↵b NA, these tests were not part of the original Canadian study.

  • Table 5.

    Comparison of reactivity in the ELISAs by animals vaccinated with B. abortus 19 in each country

    Country% for assay
    IELISA-ADRIIELISA-IAEACELISA-OCCELISA-sLPS
    ArgentinaNAd NANANA
    Canada56.30NANA97.7
    Chile78.8296.8594.44100
    Colombia86.7687.5795.5092.25
    Costa Ricab 91.8094.5893.1296.03
    Costa Ricac 95.5397.5697.5697.97
    Combineda 90.5394.5596.0896.51
    • ↵a Combined data for all the countries except Canada and Argentina.

    • ↵b Calf vaccination.

    • ↵c Adult vaccination.

    • ↵d NA, a separate vaccinated population for Argentina was not available.

  • Table 6.

    Comparison of cutoff values determined for each ELISA in each country

    Country% for assay
    IELISA-ADRIaIELISA-IAEAaCELISA-OCbCELISA-sLPSb
    Argentina67403544
    Canada46NAc NA30
    Chile16211827
    Colombia40143029
    Costa Rica70732018
    Combined41412629
    • ↵a Cutoff value is expressed as percent positivity.

    • ↵b Cutoff value is expressed as percent inhibition.

    • ↵c NA, data not available.

  • Table 7.

    Comparison of agreement among the ELISAs in each country by kappa statisticsa

    Assay and countryValue for assay
    IELISA-ADRIIELISA-IAEACELISA-OC
    IELISA-IAEA
     Costa Rica0.850
     Colombia0.994
     Chile0.963
     Argentina0.824
    CELISA-OC
     Costa Rica0.7560.796
     Colombia0.9890.989
     Chile0.9720.964
     Argentina0.8120.910
    CELISA-sLPS
     Costa Rica0.8250.7930.720
     Colombia0.9390.9390.939
     Chile0.9780.9650.981
     Argentina0.8550.9270.931
    • ↵a A kappa value of 1 is considered to indicate complete agreement.

PreviousNext
Back to top
Download PDF
Citation Tools
Enzyme Immunoassays for Serological Diagnosis of Bovine Brucellosis: A Trial in Latin America
D. Gall, A. Colling, O. Marino, E. Moreno, K. Nielsen, B. Perez, L. Samartino
Clinical and Diagnostic Laboratory Immunology Sep 1998, 5 (5) 654-661; DOI:

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print

Email

Thank you for sharing this Clinical and Vaccine Immunology article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Enzyme Immunoassays for Serological Diagnosis of Bovine Brucellosis: A Trial in Latin America
(Your Name) has forwarded a page to you from Clinical and Vaccine Immunology
(Your Name) thought you would be interested in this article in Clinical and Vaccine Immunology.
Share
Enzyme Immunoassays for Serological Diagnosis of Bovine Brucellosis: A Trial in Latin America
D. Gall, A. Colling, O. Marino, E. Moreno, K. Nielsen, B. Perez, L. Samartino
Clinical and Diagnostic Laboratory Immunology Sep 1998, 5 (5) 654-661; DOI:
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • MATERIALS AND METHODS
    • RESULTS
    • DISCUSSION
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

Related Articles

Cited By...

About

  • About CVI
  • For Librarians
  • For Advertisers
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • Submit a Manuscript to mSphere

ASM Journals

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

About ASM | Contact Us | Press Room

 

ASM is a member of

Scientific Society Publisher Alliance

Copyright © 2019 American Society for Microbiology | Privacy Policy | Website feedback

Print ISSN: 1556-6811; Online ISSN: 1556-679X