Predicting the presence and titre of rabies virus‐neutralizing antibodies from low‐volume serum samples in low‐containment facilities

Meza, D. K. , Broos, A. , Becker, D. J., Behdenna, A., Willett, B. J. , Viana, M. and Streicker, D. G. (2021) Predicting the presence and titre of rabies virus‐neutralizing antibodies from low‐volume serum samples in low‐containment facilities. Transboundary and Emerging Diseases, 68(3), pp. 1564-1576. (doi: 10.1111/tbed.13826) (PMID:32931658)

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Abstract

Serology is a core component of the surveillance and management of viral zoonoses. Virus neutralization tests are a gold standard serological diagnostic, but requirements for large volumes of serum and high biosafety containment can limit widespread use. Here, focusing on Rabies lyssavirus, a globally important zoonosis, we developed a pseudotype micro‐neutralization rapid fluorescent focus inhibition test (pmRFFIT) that overcomes these limitations. Specifically, we adapted an existing micro‐neutralization test to use a green fluorescent protein‐tagged murine leukaemia virus pseudotype in lieu of pathogenic rabies virus, reducing the need for specialized reagents for antigen detection and enabling use in low‐containment laboratories. We further used statistical models to generate rapid, quantitative predictions of the probability and titre of rabies virus‐neutralizing antibodies from microscopic imaging of neutralization outcomes. Using 47 serum samples from domestic dogs with neutralizing antibody titres estimated using the fluorescent antibody virus neutralization test (FAVN), pmRFFIT showed moderate sensitivity (78.79%) and high specificity (84.62%). Despite small conflicts, titre predictions were correlated across tests repeated on different dates both for dog samples (r = 0.93) and in a second data set of sera from wild common vampire bats (r = 0.72, N = 41), indicating repeatability. Our test uses a starting volume of 3.5 µl of serum, estimates titres from a single dilution of serum rather than requiring multiple dilutions and end point titration, and may be adapted to target neutralizing antibodies against alternative lyssavirus species. The pmRFFIT enables high‐throughput detection of rabies virus‐neutralizing antibodies in low‐biocontainment settings and is suited to studies in wild or captive animals where large serum volumes cannot be obtained.

Item Type:Articles
Additional Information:Acknowledgements: This work was funded by a Sir Henry Dale Fellowship jointly funded by the Wellcome Trust and Royal Society (102507/Z/13/Z) and a Wellcome Senior Research Fellowship (217221/Z/19/Z). Additional funding was provided by the National Science Foundation (grant: DEB-1020966). DKM was funded by the Human Frontier Science Program (grant: RGP0013/2018) and the Mexican National Council for Science and Technology (CONACYT, 334795/472296). AB was supported by the Leverhulme Trust (grant RPG-2015- 259).
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Willett, Professor Brian and Viana, Dr Mafalda and Villa Meza, Diana and Broos, Ms Alice and Behdenna, Mr Abdelkader and Streicker, Dr Daniel
Authors: Meza, D. K., Broos, A., Becker, D. J., Behdenna, A., Willett, B. J., Viana, M., and Streicker, D. G.
College/School:College of Medical Veterinary and Life Sciences > Institute of Biodiversity Animal Health and Comparative Medicine
College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Transboundary and Emerging Diseases
Publisher:Wiley
ISSN:1865-1674
ISSN (Online):1865-1682
Published Online:15 September 2020
Copyright Holders:Copyright © 2020 The Authors
First Published:First published in Transboundary and Emerging Diseases 68(3):1564-1576
Publisher Policy:Reproduced under a Creative Commons licence

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
169793Managing viral emergence at the interface of bats and livestockDaniel StreickerWellcome Trust (WELLCOTR)102507/Z/13/ZRInstitute of Biodiversity, Animal Health and Comparative Medicine
307106Epidemiology meets biotechnology: preventing viral emergence from batsDaniel StreickerWellcome Trust (WELLCOTR)217221/Z/19/ZInstitute of Biodiversity, Animal Health and Comparative Medicine
172006From observation to intervention: overcoming weak data with new approaches to complex biological problemsDaniel HaydonLeverhulme Trust (LEVERHUL)RPG-2015-259III-MRC-GU Centre for Virus Research