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      Evaluation of the comparative accuracy of the complement fixation test, Western blot and five enzyme-linked immunosorbent assays for serodiagnosis of glanders

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          Abstract

          Glanders is a zoonotic contagious disease of equids caused by Burkholderia (B.) mallei. Serodiagnosis of the disease is challenging because of false-positive and false-negative test results. The accuracy of the complement fixation test (CFT) which is prescribed for international trade by the World Organisation for Animal Health (OIE), five ELISAs and a Western blot (WB) were compared for serodiagnosis of glanders using sera from 3,000 glanders-free and 254 glanderous equids. Four ELISA tests are based on recombinant antigens (TssA, TssB, BimA and Hcp1), the IDVet ELISA is based on a semi-purified fraction of B. mallei and WB makes use of a purified LPS-containing B. mallei-antigen. Sensitivity and specificity of tests were estimated using cut-off values recommended by the test developers. The WB and all ELISAs, except BimA, were significantly more specific than the CFT. ELISAs based on TssA, TssB, and BimA antigens had significantly lower sensitivity compared to CFT while the sensitivities of the Hcp1-ELISA, the IDVet-ELISA and the WB did not differ significantly from that of the CFT. Given their comparable sensitivities and specificities, the CFT (98.0%, 96.4%), the WB (96.8%, 99.4%), the Hcp1-ELISA (95.3%, 99.6%) and the IDVet-ELISA (92.5%, 99.5%) should be further developed to meet OIE requirements.

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          Estimation of diagnostic-test sensitivity and specificity through Bayesian modeling.

          We review recent Bayesian approaches to estimation (based on cross-sectional sampling designs) of the sensitivity and specificity of one or more diagnostic tests. Our primary goal is to provide veterinary researchers with a concise presentation of the computational aspects involved in using the Bayesian framework for test evaluation. We consider estimation of diagnostic-test sensitivity and specificity in the following settings: (i) one test in one population, (ii) two conditionally independent tests in two or more populations, (iii) two correlated tests in two or more populations, and (iv) three tests in two or more populations, where two tests are correlated but jointly independent of the third test. For each scenario, we describe a Bayesian model that incorporates parameters of interest. The WinBUGS code used to fit each model, which is available at http://www.epi.ucdavis.edu/diagnos-tictests/, can be altered readily to conform to different data.
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            Validation of serological assays for diagnosis of infectious diseases.

            Assay validation is a series of the following interrelated processes: an experimental process: reagents and protocols are optimised by experimentation to detect the analyte with accuracy and precision, and to ensure repeatability and reproducibility in the assay. a relative process: its diagnostic sensitivity and diagnostic specificity are calculated relative to test results obtained from reference animal populations of known infection/exposure status. a conditional process: classification of animals in the target population as infected or uninfected is conditional upon how well the reference animal population used to validate the assay represents the population to which the assay will be applied (accurate predictions of the infection status of animals from test results and predictive values of positive and negative test results are conditional upon the estimated prevalence of disease/infection in the target population) an incremental process: confidence in the validity of an assay increases over time when use confirms that it is robust as demonstrated by accurate and precise results (the assay may also achieve increasing levels of validity as it is upgraded and extended by adding reference populations of known infection status) a continuous process: the assay remains valid only insofar as the assay continues to provide accurate and precise results as proved through statistical verification. Therefore, validation of diagnostic assays for infectious diseases does not end with a time-limited series of experiments based on a few reference samples. Rather, it is a process that also requires constant vigilance and maintenance, along with reassessment of its performance characteristics for each population of animals to which it is applied. It is certain that the current movement to develop and implement accreditation criteria for veterinary diagnostic laboratories may be of little worth unless there is some assurance that the assays conducted in such laboratories are properly validated. Fully accredited laboratories may generate highly reproducible test results, but the results may still misclassify animals as to their infection status due to an improper assay validation process. Therefore, assay validation is foundational to the core product of veterinary diagnostic laboratories--test results and their interpretation.
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              Epidemiologic issues in the validation of veterinary diagnostic tests.

              In this review, we critically discuss the objectives, methods and limitations of different approaches for the validation of diagnostic tests. We show (based on published data and our own experiences) that estimates for the diagnostic sensitivity and specificity may vary among populations and/or subpopulations of animals, conditional on the distribution of influential covariates. Additional variability in those parameter estimates may be attributable to the sampling strategy. The uncertainty about diagnostic parameters is of concern for the decision-maker in the context of clinical diagnosis or quantitative risk assessment as well as for the epidemiologist who uses test data for prevalence estimation or risk-factor studies. Examples for the calculation of diagnostic parameters are presented together with bias-avoidance strategies. We suggest guidelines for an epidemiologic approach to test validation of veterinary diagnostic tests.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: ValidationRole: Writing – original draft
                Role: ConceptualizationRole: MethodologyRole: Writing – review & editing
                Role: ConceptualizationRole: MethodologyRole: ResourcesRole: Writing – review & editing
                Role: ResourcesRole: Supervision
                Role: ConceptualizationRole: Resources
                Role: ConceptualizationRole: InvestigationRole: MethodologyRole: SoftwareRole: Writing – review & editing
                Role: InvestigationRole: Methodology
                Role: ConceptualizationRole: Methodology
                Role: ConceptualizationRole: Writing – original draft
                Role: ConceptualizationRole: InvestigationRole: Writing – review & editing
                Role: ConceptualizationRole: ResourcesRole: Writing – review & editing
                Role: ConceptualizationRole: MethodologyRole: Resources
                Role: ConceptualizationRole: MethodologyRole: SoftwareRole: Writing – review & editing
                Role: ConceptualizationRole: Funding acquisitionRole: ResourcesRole: SupervisionRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                5 April 2019
                2019
                : 14
                : 4
                : e0214963
                Affiliations
                [1 ] Institute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut (FLI), Federal Research Institute for Animal Health, Jena, Germany
                [2 ] Paris Est University, Animal Health Laboratory, EU-Reference Laboratory for Glanders, Maisons Alfort Cedex, France
                [3 ] Indian Council of Agricultural Research—National Research Centre on Equines, Hisar, India
                [4 ] Department of Clinical Medicine and Surgery, University of Agriculture Faisalabad, Faisalabad, Pakistan
                [5 ] Department of Health Management, Atlantic Veterinary College, UPEI, Prince Edward Island, Canada
                [6 ] Defence Research and Development Establishment, Microbiology Division, Gwalior, India
                [7 ] Faculty of Veterinary Medicine, Kafrelsheikh University, Kafr El-Sheikh, Egypt
                [8 ] Section of Epidemiology and Public Health, College of Veterinary and Animal Sciences, Jhang, Pakistan
                [9 ] Chaudhary Charan Singh, National Institute of Animal Health, Department of Animal Husbandry, Dairying and Fisheries, Baghpat, India
                [10 ] Institute of Epidemiology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald-Insel Riems, Germany
                Institut National de la Recherche Agronomique, FRANCE
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0001-5705-1659
                Article
                PONE-D-18-36467
                10.1371/journal.pone.0214963
                6450644
                30951554
                354bcde5-66ef-43bc-b35f-2534b6429c15
                © 2019 Elschner et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 14 January 2019
                : 23 March 2019
                Page count
                Figures: 0, Tables: 5, Pages: 12
                Funding
                Funded by: OIE and the International Horse Sports Confederation
                Award ID: Tender Ref.: AD/SR/2015/1885
                Award Recipient :
                This work was primarily funded by the International Horse Sports Confederation and World Organization of Animal Health (OIE) (Tender Ref.: AD/SR/2015/1885), grant to MCE, HS, KL, SM, IK, PM. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Organisms
                Bacteria
                Burkholderia
                Burkholderia Mallei
                Biology and Life Sciences
                Microbiology
                Medical Microbiology
                Microbial Pathogens
                Bacterial Pathogens
                Burkholderia
                Burkholderia Mallei
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Pathogens
                Microbial Pathogens
                Bacterial Pathogens
                Burkholderia
                Burkholderia Mallei
                Medicine and Health Sciences
                Infectious Diseases
                Bacterial Diseases
                Glanders
                Research and Analysis Methods
                Immunologic Techniques
                Immunoassays
                Enzyme-Linked Immunoassays
                Biology and Life Sciences
                Organisms
                Eukaryota
                Animals
                Vertebrates
                Amniotes
                Mammals
                Equines
                Horses
                Medicine and Health Sciences
                Diagnostic Medicine
                Serodiagnosis
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Serology
                Serodiagnosis
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Serology
                Biology and Life Sciences
                Biochemistry
                Proteins
                Recombinant Proteins
                People and Places
                Geographical Locations
                Europe
                Custom metadata
                All relevant data are within the manuscript and its Supporting Information files.

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                Uncategorized

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