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      Early detection of Mycobacterium avium subsp. paratuberculosis infection in cattle with multiplex-bead based immunoassays

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          Abstract

          Johne’s Disease (JD), caused by Mycobacterium avium subspecies paratuberculosis (MAP), results in significant economic loss to livestock production. The early detection of MAP infection in animals with extant serological assays has remained challenging due to the low sensitivity of commercially available ELISA tests, a fact that has hampered the development of effective JD control programs. Our recent protein microarray-based studies identified several promising candidate antigens that are immunogenic during different stages of MAP infection. To evaluate these antigens for use in diagnostic assays and reliably identify animals with MAP infection, a multiplex (Luminex ®) assay was developed using color-coded flourescent beads coupled to 6 MAP recombinant proteins and applied to screen 180 serum and 90 milk samples from cows at different stages of MAP infection including negative (NL), fecal test positive/ELISA negative (F+E-), and fecal positive/ELISA positive (F+E+). The results show that while serum antibody reactivities to each of the 6 antigens were highest in F+E+ group, antibody reactivity to three of the six antigens were identified in the F+E- group, suggesting that these three antigens are expressed and provoke antibody responses during the early infection stages with MAP. Further, antibodies against all six antigens were elevated in milk samples from both the F+E- and F+E+ groups in comparison to the NL group ( p<0.01). Taken together, the results of our investigation suggest that multiplex bead-based assays are able to reliably identify MAP infection, even during early stages when antibody responses in animals are undetectable with widely used commercial ELISA tests.

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          Most cited references36

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          Principles and practical application of the receiver-operating characteristic analysis for diagnostic tests.

          We review the principles and practical application of receiver-operating characteristic (ROC) analysis for diagnostic tests. ROC analysis can be used for diagnostic tests with outcomes measured on ordinal, interval or ratio scales. The dependence of the diagnostic sensitivity and specificity on the selected cut-off value must be considered for a full test evaluation and for test comparison. All possible combinations of sensitivity and specificity that can be achieved by changing the test's cut-off value can be summarised using a single parameter; the area under the ROC curve. The ROC technique can also be used to optimise cut-off values with regard to a given prevalence in the target population and cost ratio of false-positive and false-negative results. However, plots of optimisation parameters against the selected cut-off value provide a more-direct method for cut-off selection. Candidates for such optimisation parameters are linear combinations of sensitivity and specificity (with weights selected to reflect the decision-making situation), odds ratio, chance-corrected measures of association (e. g. kappa) and likelihood ratios. We discuss some recent developments in ROC analysis, including meta-analysis of diagnostic tests, correlated ROC curves (paired-sample design) and chance- and prevalence-corrected ROC curves.
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            Perspectives on Immunoglobulins in Colostrum and Milk

            Immunoglobulins form an important component of the immunological activity found in milk and colostrum. They are central to the immunological link that occurs when the mother transfers passive immunity to the offspring. The mechanism of transfer varies among mammalian species. Cattle provide a readily available immune rich colostrum and milk in large quantities, making those secretions important potential sources of immune products that may benefit humans. Immune milk is a term used to describe a range of products of the bovine mammary gland that have been tested against several human diseases. The use of colostrum or milk as a source of immunoglobulins, whether intended for the neonate of the species producing the secretion or for a different species, can be viewed in the context of the types of immunoglobulins in the secretion, the mechanisms by which the immunoglobulins are secreted, and the mechanisms by which the neonate or adult consuming the milk then gains immunological benefit. The stability of immunoglobulins as they undergo processing in the milk, or undergo digestion in the intestine, is an additional consideration for evaluating the value of milk immunoglobulins. This review summarizes the fundamental knowledge of immunoglobulins found in colostrum, milk, and immune milk.
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              Herd-level economic losses associated with Johne's disease on US dairy operations.

              Johne's disease ('paratuberculosis') is a chronic, infectious, wasting disease that affects dairy cattle. Estimation of its impact on herd productivity and corresponding economic loss on US dairy operations was part of the USDA National Animal Health Monitoring System's (NAHMS) 1996 national dairy study. Johne's-positive herds experience an economic loss of almost US\(100 per cow when compared to Johne's-negative herds due to reduced milk production and increased cow-replacement costs. For Johne's-positive herds that reported at least 10% of their cull cows as having clinical signs consistent with Johne's disease, economic losses were over US\) 200 per cow. These high-prevalence herds experienced reduced milk production of over 700 kg per cow, culled more cows but had lower cull-cow revenues, and had greater cow mortality than Johne's-negative herds. Averaged across all herds, Johne's disease costs the US dairy industry, in reduced productivity, US\(22 to US\) 27 per cow or US\(200 to US\) 250 million annually.
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                Author and article information

                Contributors
                Role: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: ValidationRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: ResourcesRole: ValidationRole: Writing – review & editing
                Role: Data curationRole: InvestigationRole: Methodology
                Role: Formal analysisRole: Writing – review & editing
                Role: ConceptualizationRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: ResourcesRole: Writing – review & editing
                Role: Formal analysisRole: Writing – review & editing
                Role: Writing – review & editing
                Role: Formal analysisRole: Writing – review & editing
                Role: Writing – review & editing
                Role: ConceptualizationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: ResourcesRole: VisualizationRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                19 December 2017
                2017
                : 12
                : 12
                : e0189783
                Affiliations
                [1 ] Department of Animal Science, The Pennsylvania State University, University Park, PA, United States of America
                [2 ] Huck Institutes of Life Sciences, The Pennsylvania State University, University Park, PA, United States of America
                [3 ] Department of Population Medicine and Diagnostic Sciences, Cornell University, Ithaca, NY, United States of America
                [4 ] National Animal Disease Center USDA-ARS, Ames, IA, United States of America
                [5 ] Antigen Discovery, Inc., Irvine, CA, United States of America
                Indian Institute of Technology Delhi, INDIA
                Author notes

                Competing Interests: LL, VK, and JPB are listed as inventor (s) on an issued patent application on mycobacterial diagnostics jointly owned by the University of Minnesota and the USDA and as such may be the beneficiaries of future royalties with the name and number: ("Mycobacterial diagnostics" U.S. Patent 7,867,704- January 11, 2011). Some of the technologies and proteins described may be considered for future patent applications by the Penn State University (LL, VK); USDA-ARS (JPB); Cornell University (YTG) and Antigen Discovery Inc (JJC). Joseph J. Campo is employed by Antigen Discovery, Inc. There are no additional patents, products in development or marketed products to declare. This does not alter our adherence to all the Frontiers policies on sharing data and materials, as detailed online in the guide for authors.

                Author information
                http://orcid.org/0000-0002-9648-0138
                Article
                PONE-D-17-36868
                10.1371/journal.pone.0189783
                5736219
                29261761
                e81e0195-2b61-4b0d-9905-8e2c42582add

                This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

                History
                : 13 October 2017
                : 1 December 2017
                Page count
                Figures: 4, Tables: 5, Pages: 16
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/100005825, National Institute of Food and Agriculture;
                Award ID: 2015-67015-23177
                Award Recipient :
                This work was supported by the United States Department of Agriculture, National institute of Food and Agriculture ( https://nifa.usda.gov/), 2015-67015-23177. The funder provided support in the form of salaries for author jjc, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of this author are articulated in the 'author contributions' section.
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                Biology and Life Sciences
                Nutrition
                Diet
                Beverages
                Milk
                Medicine and Health Sciences
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                All relevant data are within the paper, its Supporting Information files, and freely available at https://scholarsphere.psu.edu/concern/generic_works/vt435gf14r/.

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