Development and Validation of Confirmatory Foot-and-Mouth Disease Virus Antibody ELISAs to Identify Infected Animals in Vaccinated Populations.

Anuj Tewari,Helen Ambrose,Antonello Di Nardo,Toru Inoue,Javier Guitian,Krupali Parekh,David James Paton,Satya Parida

doi:10.3390/v13050914

Anuj Tewari, Helen Ambrose + Show 6 more

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https://doi.org/10.3390/v13050914

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Abstract

In foot-and-mouth disease (FMD)-endemic countries, vaccination is commonly used to control the disease, whilst in FMD-free countries, vaccination is considered as an option, in addition to culling the infected and in contact animals. FMD vaccines are mainly comprised of inactivated virions and stimulate protective antibodies to virus structural proteins. In contrast, infection with FMD virus leads to virus replication and additional antibody responses to viral nonstructural proteins (NSP). Therefore, antibodies against NSPs are used to differentiate infection in vaccinated animals (DIVA), in order to estimate the prevalence of infection or its absence. Another advantage of NSP antibody tests is that they detect FMD infection in the field, irrespective of the serotypes of virus in circulation. In cattle, the NSP tests that target the 3ABC polyprotein provides the highest sensitivity, detecting up to 90% of vaccinated animals that become carriers after exposure to infection, with a specificity of around 99%. Due to insufficient diagnostic sensitivity and specificity, detection of a low level of infection is difficult at the population level with a high degree of confidence. The low level of non-specific responses can be overcome by retesting samples scored positive using a second confirmatory test, which should have at least comparable sensitivity to the first test. In this study, six in-house tests were developed incorporating different NSP antigens, and validated using bovine sera from naïve animals, field cases and experimentally vaccinated and/or infected animals. In addition, two (short and long incubation) new commercial NSP tests based on 3ABC competitive blocking ELISAs (ID Screen® FMD NSP Competition, IDvet, France) were validated in this study. The two commercial ELISAs had very similar sensitivities and specificities that were not improved by lengthening the incubation period. Several of the new in-house tests had performance characteristics that were nearly as good as the commercial ELISAs. Finally, the in-house tests were evaluated for use as confirmatory tests following screening with the PrioCHECK® and ID Screen® FMDV NS commercial kits, to assess the diagnostic performance produced by a multiple testing strategy. The in-house tests could be used in series (to confirm) or in parallel (to augment) with the PrioCHECK® and IDvet® FMDV NS commercial kits, in order to improve either the specificity or sensitivity of the overall test system, although this comes at the cost of a reduction in the counterpart (sensitivity/specificity) parameter.

Highlights

Vaccination is widely used for foot-and-mouth disease (FMD) control in endemic countries
From the receiver operating characteristic (ROC) analysis, the best trade-off between specificity and sensitivity was estimated at a cut-off point of 0.5 OD, which was selected for all in-house ELISAs
Following the economic losses experienced during the UK 2001 FMD outbreak, efforts have been made to facilitate the adoption of a ‘vaccinate-to-live’ control policy, which consists of an emergency ring-vaccination within the surrounding areas of the infected premises followed by sero-surveillance to substantiate the absence of virus carrier animals and of virus circulation and to support a declaration of freedom from FMD infection [1]

Summary

Introduction

Vaccination is widely used for foot-and-mouth disease (FMD) control in endemic countries. Some countries use vaccination to maintain freedom, whilst others without FMD virus (FMDV) infection only consider vaccination as an option if the disease is introduced. This form of emergency vaccination may be followed by retention or removal of the vaccinated animals through the adoption of so-called ‘vaccination-to-live’ or ‘vaccination-to-kill’ policies, according to the urgency with which the FMD-free status is to be recovered [1]. FMD vaccination can protect animals against clinical disease and reduce or eliminate virus circulation. Until virus circulation stops, vaccinated animals can become infected with or without showing clinical disease [2]. NSP tests are used to differentiate infection in vaccinated animals (i.e., DIVA test)

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