Serological Detection of SARS-CoV-2 Antibodies in Naturally-Infected Mink and Other Experimentally-Infected Animals

Abstract

The recent emergence of SARS-CoV-2 in humans from a yet unidentified animal reservoir and the capacity of the virus to naturally infect pets, farmed animals and potentially wild animals has highlighted the need for serological surveillance tools. In this study, the luciferase immunoprecipitation systems (LIPS), employing the spike (S) and nucleocapsid proteins (N) of SARS-CoV-2, was used to examine the suitability of the assay for antibody detection in different animal species. Sera from SARS-CoV-2 naturally-infected mink (n = 77), SARS-CoV-2 experimentally-infected ferrets, fruit bats and hamsters and a rabbit vaccinated with a purified spike protein were examined for antibodies using the SARS-CoV-2 N and/or S proteins. From comparison with the known neutralization status of the serum samples, statistical analyses including calculation of the Spearman rank-order-correlation coefficient and Cohen’s kappa agreement were used to interpret the antibody results and diagnostic performance. The LIPS immunoassay robustly detected the presence of viral antibodies in naturally infected SARS-CoV-2 mink, experimentally infected ferrets, fruit bats and hamsters as well as in an immunized rabbit. For the SARS-CoV-2-LIPS-S assay, there was a good level of discrimination between the positive and negative samples for each of the five species tested with 100% agreement with the virus neutralization results. In contrast, the SARS-CoV-2-LIPS-N assay did not consistently differentiate between SARS-CoV-2 positive and negative sera. This study demonstrates the suitability of the SARS-CoV-2-LIPS-S assay for the sero-surveillance of SARS-CoV-2 infection in a range of animal species.

Evidence of antibodies against SARS-CoV-2 in wild mustelids from Brittany (France)

“…Recently, another multispecies ELISA based on the detection of antibodies against the receptor-binding domain (RBD) of SARS-CoV-2 has been developed with a specificity of 100% and sensibility of 98.3% (Wernike et al, 2020). In addition, the luciferase immunoprecipitation systems (LIPS), employing the spike, gave good results in experimentally infected ferrets (Berguido et al, 2021).…”

Variations in Cell Surface ACE2 Levels Alter Direct Binding of SARS-CoV-2 Spike Protein and Viral Infectivity: Implications for Measuring Spike Protein Interactions with Animal ACE2 Orthologs

“…It should be noted that measuring ACE2 interactions with SARS-CoV-2 may not be a perfect method for predicting a species susceptibility to infection, progression to disease, and even viral transmission. For instance, both in silico and in vitro studies have suggested that ferrets and mink ACE2 interacts poorly with S-protein [6,9,48], however these animals are readily infectible with SARS-CoV-2, often dramatically so [49][50][51][52]. As the field progresses, it will behoove us to think not only of ACE2 but other physiological differences between species which may impact SARS-CoV-2 susceptibility.…”

Development of a highly sensitive Gaussia luciferase immunoprecipitation assay for the detection of antibodies against African swine fever virus

“…Various luciferase genes have been used as the reporter gene in LIPS assays, including Renilla luciferase ( Berguido et al., 2021 ; Hachim et al., 2022 ), NanoLuciferase ( Honda et al., 2021 ), and GLuc ( Honda et al., 2021 ). GLuc from the marine copepod Gaussia princeps is naturally secreted from mammalian cells in an active form and can be released efficiently into the conditioned medium of mammalian cells ( Tannous, 2009 ).…”