Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 193, Issue 2, Pages 295-303Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2013.06.030
Keywords
Serology; Multiplex; Eidolon helvum; Emerging diseases; Microsphere binding assay
Funding
- University of Cambridge, Institute of Zoology
- Australian Animal Health Laboratory
- Charles Slater Trust
- Zebra Foundation for Veterinary Zoological Education provided travel
- Biotechnology and Biological Sciences Research Council [BB/I012192/1]
- Wellcome Trust
- Research and Policy for Infectious Disease Dynamics (RAPIDD)
- Science and Technology Directorate
- Department of Homeland Security
- Fogarty International Center, USA
- Department for Environment
- Food and Rural Affairs (Defra) [SEV3500]
- Royal Society Wolfson Research Merit
- Alborada Trust
- BBSRC [BB/I012192/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/I012192/1] Funding Source: researchfish
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Reservoir hosts of novel pathogens are often identified or suspected as such on the basis of serological assay results, prior to the isolation of the pathogen itself. Serological assays might therefore be used outside of their original, validated scope in order to infer seroprevalences in reservoir host populations, until such time that specific diagnostic assays can be developed. This is particularly the case in wildlife disease research. The absence of positive and negative control samples and gold standard diagnostic assays presents challenges in determining an appropriate threshold, or 'cutoff', for the assay that enables differentiation between seronegative and seropositive individuals. Here, multiple methods were explored to determine an appropriate cutoff for a multiplexed microsphere assay that is used to detect henipavirus antibody binding in fruit bat plasma. These methods included calculating multiples of `negative' control assay values, receiver operating characteristic curve analyses, and Bayesian mixture models to assess the distribution of assay outputs for classifying seropositive and seronegative individuals within different age classes. As for any diagnostic assay, the most appropriate cutoff determination method and value selected must be made according to the aims of the study. This study is presented as an example for others where reference samples, and assays that have been characterised previously, are absent. (C) 2013 The Authors. Published by Elsevier B.V. All rights reserved.
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