Development and Evaluation of a Duplex Lateral Flow Assay for the Detection and Differentiation between Rabbit Haemorrhagic Disease Virus Lagovirus europaeus/GI.1 and /GI.2.

Alba Fresco-Taboada,Paloma Rueda,Elena Soria,Cécile Guillou-Cloarec,Juan Bárcena,Mercedes Montón,Ghislaine Le Gall-Reculé,Istar Tapia,Esther Blanco

doi:10.3390/biology11030401

Abstract

Simple SummaryRabbit Haemorrhagic Disease is caused by a virus that affects the liver, the spleen and the lungs of rabbits, causing hepatitis, splenomegaly and haemorrhages. A new genotype of the virus was first reported in France in 2010 and has spread globally since then, replacing most of the circulating former viruses in many countries. The detection of the virus and the differentiation of both genotypes is of crucial importance for disease surveillance. In this article, a rapid test for antigen detection is described and evaluated, providing the first description of a quick and easy-to-use test that allows for the simultaneous detection and differentiation of the genotypes. A total of 136 samples, rabbit liver samples and liver exudates (liquid collected after freeze–thawing) classified as infected and non-infected, were analysed, with good results. These data confirm that the developed rapid test can be used as a reliable diagnostic test for disease surveillance, especially in farms and the field.Rabbit Haemorrhagic Disease Virus 2 (RHDV2, recently named Lagovirus europaeus/GI.2) was first reported in France in 2010 and has spread globally since then, replacing most of the circulating former RHDV (genotype GI.1) in many countries. The detection and differentiation of both genotypes is of crucial importance for the surveillance of the disease. In this article, a duplex lateral flow assay (LFA) for antigen detection is described and evaluated, providing the first description of a quick and easy-to-use test that allows for the simultaneous detection and differentiation of RHDV genotypes GI.1 and GI.2. A panel of GI.1- or GI.2-infected and non-infected rabbit liver samples and liver exudates (136 samples) was analysed, obtaining a total sensitivity of 94.4% and specificity of 100%. These data confirm that the developed duplex LFA can be used as a reliable diagnostic test for RHD surveillance, especially in farms and the field.

Highlights

Considering all the liver samples, the results indicated a specificity of 100% and a sensitivity of 93.8% compared with the room temperature (RT)-PCR results
Rabbit Haemorrhagic Disease is a deadly disease for rabbits that belongs to the OIE-list
The other matrices could be very useful in disease surveillance in wild and farming rabbits, as the sample is simpler to extract in live and dead animals that could be encountered in the field. These results show that the newly developed duplex lateral flow assay is a suitable tool for the differential diagnosis of GI.1 and GI.2 outbreaks in a single strip

Summary

Introduction

Rabbit Haemorrhagic Disease Virus (RHDV), a member of the genus Lagovirus Caliciviridae), causes a fatal disease in domestic and wild European rabbits (Oryctolagus cuniculus) and was first reported in China in 1984. Since it has been reported in over forty countries in the five continents, being endemic in most parts of the world where. In 2010, a new pathogenic lagovirus, phylogenetically distinct from any previously described pathogenic and benign members of the genus, was identified in France [3]. It presents with a unique antigenic profile [4]

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