RNA in-situ hybridization for pathology-based diagnosis of feline infectious peritonitis (FIP): current diagnostics for FIP and comparison to the current gold standard

Sweet, Arjun; André, Nicole M.; Whittaker, Gary

doi:10.32388/nun8kb

Cited by 3 publications

(2 citation statements)

References 33 publications

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“…Mapping of the nucleotides in each branch of the loops shows that each contains mutations in codons from amino acids within the FCS (P6, P3', P4' in Figure 3). To examine the possible tissue distribution of FCoV-1, samples from necropsy were examined by histopathology, using a newly developed RNA-in-situ hybridization technique (Sweet et al, 2022), with the goal of improving sensitivity in the possible sites of viral persistence. Using this technique, FCoV RNA was detected in isolated cells in the heart, and in epithelial cells of the small intestine (jejunum) (Figure 5).…”

Section: Resultsmentioning

confidence: 99%

“…A probe targeting the RdRp gene of FCoV was used to detect viral RNA (RNAscope ® 2.5 VS Probe V-FIPV-ORF1a1b, Advanced Cell Diagnostics) using ISH (Sweet et al, 2022). The ISH process was carried by the AHDC using the automated staining platform Ventana Discovery Ultra (Roche Tissue Diagnostics) using the Discovery kits for mRNA Sample Prep, mRNA Red Probe Amplification and mRNA Red Detection and the Ventana Hematoxylin and Ventana Bluing Reagents for counterstaining.…”

Section: Rna-based In Situ Hybridization (Ish)mentioning

confidence: 99%

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Intra-host variation in the spike S1/S2 region of a feline coronavirus type-1 in a cat with persistent infection

Olarte‐Castillo

Licitra

André

et al. 2023

Preprint

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Feline coronavirus type 1 (FCoV-1) is widely known for causing feline infectious peritonitis (FIP), a systemic infection that is often fatal, with the virus known as the FIPV biotype. However, subclinical disease also occurs, in which cats may not show signs and intermittently shed the virus, including in feces, possibly for long periods of time. This virus is known as the FECV biotype. Progression of FECV to FIPV has been linked to several genomic changes, however a specific region of the viral spike protein at the interface of the spike S1 and S2 domains has been especially implicated. In this study, we followed a cat (#576) for six years from 2017, at which time FCoV-1 was detected in feces and conjunctival swabs, until 2022, when the animal was euthanized based on a diagnosis of alimentary small cell lymphoma. Over this time period, the cat was clinically diagnosed with inflammatory bowel disease and chronic rhinitis, and cardiac problems were also suspected. Using hybridization capture targeting the spike (S) gene of FCoV followed by next-generation sequencing, we screened 27 clinical samples. We detected FCoV-1 in 4 samples taken in 2017 (intestine and nasal tissue, feces, and conjunctiva), and 3 samples taken in 2022 (feces, and intestinal and heart tissue), but not in fecal samples taken in 2019 and 2020. Next, we focused on the S1/S2 region within S, which contains the furin cleavage site (FCS), a key regulator of viral transmission and pathogenesis. We show that the FCoV-1 variants obtained from feces in 2017 and 2022 were identical, while the ones from conjunctiva (2017), heart (2022), and intestine (2017 and 2022) were distinct. Sequence comparison of all the variants obtained showed that most of the non-synonymous changes in the S1/S2 region occur within the FCS. In the heart, we found two variants that differed by a single nucleotide, resulting in distinct FCS motifs that differ in one amino acid. It is predicted that one of these FCS motifs will down-regulate spike cleavability. The variant from the conjunctiva (2017) had a 6-nucleotide in-frame insertion that resulted in a longer and more exposed S1/S2 loop, which is predicted to be more accessible to the furin protease. Our studies indicate that FCoV-1 can independently persist in the gastrointestinal tract and heart of a cat over a long period of time without evidence of typical FIP signs, with intermittent viral shedding from the gastrointestinal and respiratory tracts.

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Section: Resultsmentioning

confidence: 99%

Section: Rna-based In Situ Hybridization (Ish)mentioning

confidence: 99%

Intra-host variation in the spike S1/S2 region of a feline coronavirus type-1 in a cat with persistent infection

Olarte‐Castillo

Licitra

André

et al. 2023

Preprint

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Persistent feline coronavirus infection in a cat with cardiac and gastrointestinal signs

Licitra,

Olarte-Castillo,

Whit-taker

2024

Companion Animal

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Background Feline coronavirus infection causes feline infectious peritonitis in a subset of cats, but can also result in persistent infection. The tissue reservoirs of feline coronavirus and the role of viral persistence in pathogenesis are poorly understood. Aims This study aimed to identify sites of feline coronavirus persistence in a naturally infected cat, identify disease correlates and characterise within-host viral evolution. Methods The study followed a 5-year-old Bengal cat for 6 years and collected non-invasive samples, including faeces and conjunctival, oropharyngeal and saliva swabs. At 11-years-old, the patient was euthanised as a result of respiratory distress, and tissue samples were collected. The authors used hybridisation capture and next-generation sequencing methodologies focused on the feline coronavirus S gene, along with RNA in-situ hybridisation. Results During the study, the patient was diagnosed with inflammatory bowel disease, alimentary small cell lymphoma, chronic rhinitis and mitral valve regurgitation. Feline coronavirus was detected in the nasal cavity, intestine, faeces and conjunctiva in 2017, and in the intestine, faeces and heart in 2022. Sequence analysis showed that the virus adapted to tissue reservoirs over time. Conclusions This study identifies potential feline coronavirus reservoirs. The relationship of persistent feline coronavirus infection to chronic conditions warrants further investigation.

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An Exploratory Bioinformatic Investigation of Cats’ Susceptibility to Coronavirus-Deriving Epitopes

Buonocore,

De Biase,

Sorrentino

et al. 2024

Life

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Coronaviruses are highly transmissible and pathogenic viruses for humans and animals. The vast quantity of information collected about SARS-CoV-2 during the pandemic helped to unveil details of the mechanisms behind the infection, which are still largely elusive. Recent research demonstrated that different class I/II human leukocyte antigen (HLA) alleles might define an individual susceptibility to SARS-CoV-2 spreading, contributing to the differences in the distribution of the infection through different populations; additional studies suggested that the homolog of the HLA in cats, the feline leukocyte antigen (FLA), plays a pivotal role in the transmission of viruses. With these premises, this study aimed to exploit a bioinformatic approach for the prediction of the transmissibility potential of two distinct feline coronaviruses (FCoVs) in domestic cats (feline enteric coronavirus (FeCV) and feline infectious peritonitis virus (FIPV)) using SARS-CoV-2 as the reference model. We performed an epitope mapping of nonapeptides deriving from SARS-CoV-2, FeCV, and FIPV glycoproteins and predicted their affinities for different alleles included in the three main loci in class I FLAs (E, H, and K). The predicted complexes with the most promising affinities were then subjected to molecular docking and molecular dynamics simulations to provide insights into the stability and binding energies in the cleft. Results showed the FLA proteins encoded by alleles in the FLA-I H (H*00501 and H*00401) and E (E*01001 and E*00701) loci are largely responsive to several epitopes deriving from replicase and spike proteins of the analyzed coronaviruses. The analysis of the most affine epitope sequences resulting from the prediction can stimulate the development of anti-FCoV immunomodulatory strategies based on peptide drugs.

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RNA in-situ hybridization for pathology-based diagnosis of feline infectious peritonitis (FIP): current diagnostics for FIP and comparison to the current gold standard

Cited by 3 publications

References 33 publications

Intra-host variation in the spike S1/S2 region of a feline coronavirus type-1 in a cat with persistent infection

Intra-host variation in the spike S1/S2 region of a feline coronavirus type-1 in a cat with persistent infection

Persistent feline coronavirus infection in a cat with cardiac and gastrointestinal signs

An Exploratory Bioinformatic Investigation of Cats’ Susceptibility to Coronavirus-Deriving Epitopes

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