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A mRNA PCR for the diagnosis of feline infectious peritonitis
Fermin A Simons1, Harry Vennema, Jaime E Rofina
1Institute of Virology, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands. f.simmons1@janbe.jnj.com
Abstract:
A reverse transcriptase polymerase chain reaction (RT-PCR) for the detection of feline coronavirus (FCoV) messenger RNA in peripheral blood mononuclear cells (PBMCs) is described. The assay is evaluated as a diagnostic test for feline infectious peritonitis (FIP). It is based on a well-documented key event in the development of FIP: the replication of virulent FCoV mutants in monocytes/macrophages. To detect most feline coronavirus field strains, the test was designed to amplify subgenomic mRNA of the highly conserved M gene. The test was applied to 1075 feline blood samples (424 from healthy, 651 from sick cats suspected of FIP) and returned 46% of the diseased cats as positive for feline coronavirus mRNA in their peripheral blood cells; of the healthy cats, 5% tested positive. Of a group of 81 animals in which FIP had been confirmed by post-mortem examination, 75 (93%) tested positive, whereas 17 cats with different pathologies (non-FIP cases) all tested negative. In view of the low rate of false-positive results (high specificity) the mRNA RT-PCR may be a valuable addition to the diagnostic arsenal for FIP.
Insights
A new RT-PCR test detects feline coronavirus mRNA in blood cells, aiding in feline infectious peritonitis diagnosis. This feline coronavirus test shows high accuracy in sick cats, offering a valuable diagnostic tool.
Area of Science:
- Veterinary Virology
- Molecular Diagnostics
- Feline Infectious Diseases
Background:
- Feline infectious peritonitis (FIP) is a severe disease caused by feline coronavirus (FCoV).
- Current diagnostic methods for FIP can be challenging, necessitating improved detection techniques.
- FIP development involves virulent FCoV replication in monocytes/macrophages.
Purpose of the Study:
- To develop and evaluate a reverse transcriptase polymerase chain reaction (RT-PCR) assay for detecting FCoV mRNA.
- To assess the utility of this RT-PCR assay as a diagnostic tool for feline infectious peritonitis (FIP).
Main Methods:
- Developed an RT-PCR assay targeting subgenomic mRNA of the conserved M gene of FCoV.
- Applied the assay to peripheral blood mononuclear cells (PBMCs) from 1075 cats (424 healthy, 651 suspected FIP).
- Validated assay performance on a cohort of 81 FIP-confirmed cases and 17 non-FIP cases.
Main Results:
- The RT-PCR assay detected FCoV mRNA in 46% of sick cats and 5% of healthy cats.
- Confirmed FIP cases showed a high positive rate (93%), while non-FIP cases were all negative.
- The assay demonstrated high specificity, indicating a low rate of false positives.
Conclusions:
- The feline coronavirus mRNA RT-PCR assay is a sensitive and specific diagnostic tool for FIP.
- This molecular assay can be a valuable addition to the diagnostic arsenal for feline infectious peritonitis.
- The test's ability to detect FCoV mRNA in PBMCs offers a promising approach for FIP diagnosis.
