@article{5e8170fc071a4d8c8cdb47ed4982faed,
title = "Ultrasensitive point-of-care immunoassay for secreted glycoprotein detects Ebola infection earlier than PCR",
abstract = "Ebola virus (EBOV) hemorrhagic fever outbreaks have been challenging to deter due to the lack of health care infrastructure in disease-endemic countries and a corresponding inability to diagnose and contain the disease at an early stage. EBOV vaccines and therapies have improved disease outcomes, but the advent of an affordable, easily accessed, mass-produced rapid diagnostic test (RDT) that matches the performance of more resource-intensive polymerase chain reaction (PCR) assays would be invaluable in containing future outbreaks. Here, we developed and demonstrated the performance of a new ultrasensitive point-of-care immunoassay, the EBOV D4 assay, which targets the secreted glycoprotein of EBOV. The EBOV D4 assay is 1000-fold more sensitive than the U.S. Food and Drug Administration approved RDTs and detected EBOV infection earlier than PCR in a standard nonhuman primate model. The EBOV D4 assay is suitable for low-resource settings and may facilitate earlier detection, containment, and treatment during outbreaks of the disease.",
author = "Fontes, {Cassio M.} and Lipes, {Barbara D.} and Jason Liu and Agans, {Krystle N.} and Aiwei Yan and Patricia Shi and Cruz, {Daniela F.} and Garrett Kelly and Luginbuhl, {Kelli M.} and Joh, {Daniel Y.} and Foster, {Stephanie L.} and Jacob Heggestad and Angus Hucknall and Mikkelsen, {Maiken H.} and Pieper, {Carl F.} and Horstmeyer, {Roarke W.} and Geisbert, {Thomas W.} and Gunn, {Michael D.} and Ashutosh Chilkoti",
note = "Funding Information: Acknowledgments: We thank T. Hoenen at Rocky Mountain Laboratories for providing EBOV sGP (Mayinga) expression vector. We also thank B. Watts at the Duke University Vaccine Institute for the assistance with SPR and R. Gordon for the GenePix usage. Funding: The authors acknowledge funding from the NIH grant R01AI150888 to A.C. and U.S. Special Operations Command (contract number W81XWH-16-C-0219) to A.C. Partial support for some filovirus samples was provided by the Department of Health and Human Services, NIH grant U19AI142785 to T.W.G. and UC7AI094660 for BSL-4 operations support of the GNL. Opinions, interpretations, conclusions, and recommendations are those of the authors and are not necessarily endorsed by the University of Texas Medical Branch or the NIH. C.M.F. also acknowledges financial support from the National Council for the Improvement of Higher Education (CAPES) and the Science without Borders project. Author contributions: C.M.F., B.D.L., and J.L. equally contributed to this project and were responsible for conceptualization, investigation, and writing of the manuscript. C.M.F. conceptualized the Ab pairing strategy and was responsible for developing and validating the EBOV D4 assay. C.M.F. was also responsible for integrating the EBOV D4 assay with the D4Scope. J.L. conceptualized and developed the D4Scope and dedicated control software. B.D.L. identified sGP as an EBOV diagnostic target, oversaw production of recombinant sGP proteins, and constructed the Ab library. B.D.L., A.Y., and P.S. identified and characterized Abs. K.N.A. was responsible for animal studies in BSL-4. D.F.C., G.K., K.M.L., D.Y.J., and J.H. were responsible for conceptualization, investigation, and manuscript writing. S.L.F. was responsible for Western blot analysis with samples from nonhuman primates in BSL-4. A.H. was responsible for assay and detector conceptualization. M.H.M. was responsible for assay conceptualization and manuscript writing. C.F.P. was responsible for statistical analysis and manuscript writing. R.W.H. was responsible for detector conceptualization and manuscript writing. T.W.G. was responsible for overseeing animal studies in BSL-4. M.D.G. initiated the Ebola diagnostic project and supervised all aspects of Ab development, engineering, and manuscript writing. A.C. was responsible for conceptualizing and overseeing the study and writing and editing the manuscript. Competing interests: The underlying technology of the D4 was developed by A.H. and A.C. and acquired by Immucor Inc. (Norcross, GA) in 2014. The other authors declare that they have no competing financial interests. Data and materials availability: All data associated with this study are present in the paper or the Supplementary Materials. Computer code that controls the D4Scope has been uploaded to Zenodo under DOI 10.5281/ zenodo.4536655. Publisher Copyright: {\textcopyright} 2021 American Association for the Advancement of Science. All rights reserved.",
year = "2021",
month = apr,
day = "7",
doi = "10.1126/scitranslmed.abd9696",
language = "English (US)",
volume = "13",
journal = "Science Translational Medicine",
issn = "1946-6234",
publisher = "American Association for the Advancement of Science",
number = "588",
}