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. 2020 Aug;22(8):791-799.
doi: 10.1177/1098612X19886671. Epub 2019 Nov 15.

Feline coronavirus with and without spike gene mutations detected by real-time RT-PCRs in cats with feline infectious peritonitis

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Feline coronavirus with and without spike gene mutations detected by real-time RT-PCRs in cats with feline infectious peritonitis

Laura Emmler et al. J Feline Med Surg. 2020 Aug.

Abstract

Objectives: Feline infectious peritonitis (FIP) emerges when feline coronaviruses (FCoVs) mutate within their host to a highly virulent biotype and the immune response is not able to control the infection. FCoV spike (S) gene mutations are considered to contribute to the change in virulence by enabling FCoV infection of and replication in macrophages. This study investigated the presence of FCoV with and without S gene mutations in cats with FIP using two different real-time RT-PCRs on different samples obtained under clinical conditions.

Methods: Fine-needle aspirates (FNAs) and incisional biopsies (IBs) of popliteal and mesenteric lymph nodes, liver, spleen, omentum and kidneys (each n = 20), EDTA blood (n = 13), buffy coat smears (n = 13), serum (n = 11), effusion (n = 14), cerebrospinal fluid (n = 16), aqueous humour (n = 20) and peritoneal lavage (n = 6) were obtained from 20 cats with FIP diagnosed by immunohistochemistry. Samples were examined by RT-PCR targeting the FCoV 7b gene, detecting all FCoV, and S gene mutation RT-PCR targeting mutations in nucleotides 23531 and 23537. The prevalence of FCoV detected in each sample type was calculated.

Results: In 20/20 cats, FCoV with S gene mutations was present in at least one sample, but there was variation in which sample was positive. FCoV with mutations in the S gene was most frequently found in effusion (64%, 95% confidence interval [CI] 39-89), followed by spleen, omentum and kidney IBs (50%, 95% CI 28-72), mesenteric lymph node IBs and FNAs (45%, 95% CI 23-67), and FNAs of spleen and liver and liver IBs (40%, 95% CI 19-62).

Conclusions and relevance: In these 20 cats with FIP, FCoVs with S gene mutations were found in every cat in at least one tissue or fluid sample. This highlights the association between mutated S gene and systemic FCoV spread. Examining a combination of different samples increased the probability of finding FCoV with the mutated S gene.

Keywords: FCoV; FIP; IHC; RT-PCR; S gene; immunohistochemistry.

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Conflict of interest statement

Christian Leutenegger was Director of Molecular Diagnostics at IDEXX Laboratories, Sacramento. Hans-Joerg Balzer is Head of Molecular Diagnostics at IDEXX Laboratories, Ludwigsburg. Nikola Pantchev is employed at IDEXX Laboratories, Ludwigsburg. The RT-PCR used in this study was performed by IDEXX Ludwigsburg. However, IDEXX was not involved in study design, in collection and interpretation of data, or in the decision to submit the manuscript for publication. There is no commercial conflict of interest. The study solely served scientific purposes. The authors declare no competing interests.

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