Feline Coronavirus Antibodies Research Articles

Serological Investigation on the Presence of Feline Coronavirus (FCoV) and Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in Domestic Cats Living with COVID-19 Positive Owners in the UAE, 2022.

Feline coronavirus (FCoV) is widely circulating among domestic cats (Felis catus). The zoonotic origin of the emerged severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and the biological characteristics of CoVs, including the ability to cross interspecies barriers, facilitate its emergence in different animals, including cats' populations. The current study is the first to report the serological investigation on the presence of FCoV and SARS-CoV-2 in domestic cats living with COVID-19-positive owners in the UAE. A total of 83 sera were collected from domestic cats living with COVID-19-positive owners (by RT-qPCR). The cats were sampled during the period between February and May 2022 in Al-Ain and Abu Dhabi Cities, UAE. Detection of FCoV and SARS-CoV-2 was carried out by enzyme-linked immunosorbent assay (ELISA). FCoV antibodies were detected in 54 samples (65%). The frequencies of FCoV were significantly higher in purebred cats (48%; 40/83) and in the cat group with outdoor access (49.4%; 41/83). SARS-CoV-2 seroprevalence in collected sera revealed 8 samples (9.6%) with positive results. Four samples (4.8%) showed positive results for both FCoV and SARS-CoV-2 antibodies. In conclusion, FCoV is widely circulating within cats' populations involved in the study. The antibodies for SARS-CoV-2 were detected in cats' populations but at a low prevalence rate. COVID-19-positive people should avoid close contact with their cats. Future serological testing of large cats' populations is crucial for providing a good understanding of COVID-19 dynamics in cats.

Alpha-1 Acid Glycoprotein Reduction Differentiated Recovery from Remission in a Small Cohort of Cats Treated for Feline Infectious Peritonitis.

Feline infectious peritonitis (FIP) is a systemic immune-mediated inflammatory perivasculitis that occurs in a minority of cats infected with feline coronavirus (FCoV). Various therapies have been employed to treat this condition, which was previously usually fatal, though no parameters for differentiating FIP recovery from remission have been defined to enable clinicians to decide when it is safe to discontinue treatment. This retrospective observational study shows that a consistent reduction of the acute phase protein alpha-1 acid glycoprotein (AGP) to within normal limits (WNL, i.e., 500 μg/mL or below), as opposed to duration of survival, distinguishes recovery from remission. Forty-two cats were diagnosed with FIP: 75% (12/16) of effusive and 54% (14/26) of non-effusive FIP cases recovered. Presenting with the effusive or non-effusive form did not affect whether or not a cat fully recovered (p = 0.2). AGP consistently reduced to WNL in 26 recovered cats but remained elevated in 16 cats in remission, dipping to normal once in two of the latter. Anaemia was present in 77% (23/30) of the cats and resolved more quickly than AGP in six recovered cats. The presence of anaemia did not affect the cat’s chances of recovery (p = 0.1). Lymphopenia was observed in 43% (16/37) of the cats and reversed in nine recovered cats but did not reverse in seven lymphopenic cats in the remission group. Fewer recovered cats (9/24: 37%) than remission cats (7/13: 54%) were lymphopenic, but the difference was not statistically different (p = 0.5). Hyperglobulinaemia was slower than AGP to return to WNL in the recovered cats. FCoV antibody titre was high in all 42 cats at the outset. It decreased significantly in 7 recovered cats but too slowly to be a useful parameter to determine discontinuation of antiviral treatments. Conclusion: a sustained return to normal levels of AGP was the most rapid and consistent indicator for differentiating recovery from remission following treatment for FIP. This study provides a useful model for differentiating recovery from chronic coronavirus disease using acute phase protein monitoring.

Development of an Indirect ELISA Based on Spike Protein to Detect Antibodies against Feline Coronavirus.

Feline coronavirus (FCoV) is a pathogenic virus commonly found in cats that causes a benign enteric illness and fatal systemic disease, feline infectious peritonitis. The development of serological diagnostic tools for FCoV is helpful for clinical diagnosis and epidemiological investigation. Therefore, this study aimed to develop an indirect enzyme-linked immunosorbent assay (iELISA) to detect antibodies against FCoV using histidine-tagged recombinant spike protein. FCoV S protein (1127–1400 aa) was expressed and used as an antigen to establish an ELISA. Mice and rabbits immunized with the protein produced antibodies that were recognized and bound to the protein. The intra-assay coefficient of variation (CV) was 1.15–5.04% and the inter-assay CV was 4.28–15.13%, suggesting an acceptable repeatability. iELISA did not cross-react with antisera against other feline viruses. The receiver operating characteristic curve analysis revealed an 86.7% sensitivity and 93.3% specificity for iELISA. Serum samples (n = 107) were tested for anti-FCoV antibodies, and 70.09% of samples were positive for antibodies against FCoV. The iELISA developed in our study can be used to measure serum FCoV antibodies due to its acceptable repeatability, sensitivity, and specificity. Additionally, field sample analysis data demonstrated that FCoV is highly prevalent in cat populations in Fujian province, China.

Correlation of Feline Coronavirus Shedding in Feces with Coronavirus Antibody Titer.

Background: Feline coronavirus (FCoV) infection is ubiquitous in multi-cat households. Responsible for the continuous presence are cats that are chronically shedding a high load of FCoV. The aim of the study was to determine a possible correlation between FCoV antibody titer and frequency and load of fecal FCoV shedding in cats from catteries. Methods: Four fecal samples from each of 82 cats originating from 19 German catteries were examined for FCoV viral loads by quantitative reverse transcriptase polymerase chain reaction (RT-qPCR). Additionally, antibody titers were determined by an immunofluorescence assay. Results: Cats with antibodies were more likely to be FCoV shedders than non-shedders, and there was a weak positive correlation between antibody titer and mean fecal virus load (Spearman r = 0.2984; p = 0.0072). Antibody titers were significantly higher if cats shed FCoV more frequently throughout the study period (p = 0.0063). When analyzing only FCoV shedders, cats that were RT-qPCR-positive in all four samples had significantly higher antibody titers (p = 0.0014) and significantly higher mean fecal virus loads (p = 0.0475) than cats that were RT-qPCR-positive in only one, two, or three samples. Conclusions: The cats’ antibody titers correlate with the likelihood and frequency of FCoV shedding and fecal virus load. Chronic shedders have higher antibody titers and shed more virus. This knowledge is important for the management of FCoV infections in multi-cat environments, but the results indicate that antibody measurement cannot replace fecal RT-qPCR.

A comparative study of techniques used for the diagnosis of effusive feline infectious peritonitis

Feline infectious peritonitis (FIP) is a fatal disease caused by feline infectious peritonitis virus (FIPV). At present, neither a licensed treatment nor an accurate ante-mortem diagnosis are available. In the present study, three available tests were evaluated for their diagnostic power on effusion samples. High feline coronavirus antibody titers, measured with an immunoperoxidase monolayer assay (IPMA), were correlated with FIP but its low specificity precluded a reliable diagnosis. The in-house 5’ reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) provided a much better specificity and high sensitivity. Given the low sensitivity of immunofluorescence staining (IF) of effusive cells, the RT-qPCR alone or in combination with IPMA represents a good alternative for IF. In the majority of the effusion samples from FIP positive animals, Sanger sequencing of the open reading frame encoding the spike protein (ORF S) revealed not only mutations that were previously associated with FIP (M1058L, S1060A, I1106T and D1108Y/E/G) but also two new, closely related mutations (T1112S/N).

Feline coronavirus antibody titer in cerebrospinal fluid from cats with neurological signs.

To investigate the utility of cerebrospinal fluid (CSF) anti-feline coronavirus (FCoV) antibody test for diagnosis of feline infectious peritonitis (FIP), the antibody titers were tested in CSF and sera from 271 FIP-suspected neurological cats. CSF antibody was detected in 28 cats, which were divided into 2 groups; 15 with CSF titer of 1:80 or lower and 13 with CSF titer of 1:640 or higher. In the latter group, reciprocal serum titer/reciprocal CSF titer was 8 or lower, which is extremely lower than normal range (256-2048), and FCoV RNA was detected in all of 11 CSF samples assayed by RT-PCR. Our findings indicate that CSF titer of 1:640 or higher may be served as a candidate for the index for diagnosing FIP.

Discrepancies between feline coronavirus antibody and nucleic acid detection in effusions of cats with suspected feline infectious peritonitis

Intra-vitam diagnosis of feline infectious peritonitis (FIP) is a challenge for veterinary diagnosticians, since there are no highly specific and sensitive assays currently available. With the aim to contribute to fill this diagnostic gap, a total of 61 effusions from cats with suspected effusive FIP were collected intra-vitam for detection of feline coronavirus (FCoV) antibodies and RNA by means of indirect immunofluorescence (IIF) assay and real-time RT-PCR (qRT-PCR), respectively. In 5 effusions there was no evidence for either FCoV RNA or antibodies, 51 and 52 specimens tested positive by IIF and qRT-PCR, respectively, although antibody titres≥1:1600, which are considered highly suggestive of FIP, were detected only in 37 effusions. Three samples with high antibody levels tested negative by qRT-PCR, whereas 18 qRT-PCR positive effusions contained no or low-titre antibodies. qRT-PCR positive samples with low antibody titres mostly contained low FCoV RNA loads, although the highest antibody titres were detected in effusions with CT values>30. In conclusion, combining the two methods, i.e., antibody and RNA detection would help improving the intra-vitam diagnosis of effusive FIP.

An eight-year epidemiologic study based on baculovirus-expressed type-specific spike proteins for the differentiation of type I and II feline coronavirus infections.

BackgroundFeline infectious peritonitis (FIP) is a fatal disease caused by feline coronavirus (FCoV). FCoVs are divided into two serotypes with markedly different infection rates among cat populations around the world. A baculovirus-expressed type-specific domain of the spike proteins of FCoV was used to survey the infection of the two viruses over the past eight years in Taiwan.ResultsAn immunofluorescence assay based on cells infected with the recombinant viruses that was capable of distinguishing between the two types of viral infection was established. A total of 833 cases from a teaching hospital was surveyed for prevalence of different FCoV infections. Infection of the type I FCoV was dominant, with a seropositive rate of 70.4%, whereas 3.5% of cats were infected with the type II FCoV. In most cases, results derived from serotyping and genotyping were highly agreeable. However, 16.7% (4/24) FIP cats and 9.8% (6/61) clinically healthy cats were found to possess antibodies against both viruses. Moreover, most of the cats (84.6%, 22/26) infected with a genotypic untypable virus bearing a type I FCoV antibody.ConclusionA relatively simple serotyping method to distinguish between two types of FCoV infection was developed. Based on this method, two types of FCoV infection in Taiwan was first carried out. Type I FCoV was found to be predominant compared with type II virus. Results derived from serotyping and genotyping support our current understanding of evolution of disease-related FCoV and transmission of FIP.

Detection of Ascitic Feline Coronavirus RNA from Cats with Clinically Suspected Feline Infectious Peritonitis

ABSTRACTAscitic feline coronavirus (FCoV) RNA was examined in 854 cats with suspected feline infectious peritonitis (FIP) by RT-PCR. The positivity was significantly higher in purebreds (62.2%) than in crossbreds (34.8%) (P<0.0001). Among purebreds, the positivities in the Norwegian forest cat (92.3%) and Scottish fold (77.6%) were significantly higher than the average of purebreds (P=0.0274 and 0.0251, respectively). The positivity was significantly higher in males (51.5%) than in females (35.7%) (P<0.0001), whereas no gender difference has generally been noted in FCoV antibody prevalence, indicating that FIP more frequently develops in males among FCoV-infected cats. Genotyping was performed for 377 gene-positive specimens. Type I (83.3%) was far more predominantly detected than type II (10.6%) (P<0.0001), similar to previous serological and genetic surveys.

Samples with high virus load cause a trend toward lower signal in feline coronavirus antibody tests.

Measurement of feline coronavirus (FCoV) antibody titres is utilised mainly for diagnosing feline infectious peritonitis (FIP) and for quarantine purposes. However, occasional samples show a falsely low or negative FCoV antibody test. We tested the hypothesis that such results are due to virus in the sample binding antibody and rendering it unavailable to antigen in the test. Thirteen effusions, one plasma and three undefined samples from cats with FIP, which gave unexpectedly low FCoV antibody titres, were examined by real-time reverse transcriptase polymerase chain reaction (RT-PCR). Increasing amounts of virus correlated with lower signals in indirect immunoflourescent, enzyme-linked immunosorbent asssay and rapid immunomigration antibody tests. However, five samples were negative by RT-PCR, so the presence of virus alone may not explain all cases of false-negative FCoV antibody tests, although it is a possible explanation in 71% of discordant samples. We conclude that falsely low or negative FCoV antibody tests can occur in samples rich in virus.

Virological Survey in free-ranging wildcats (Felis silvestris) and feral domestic cats in Portugal

To determine the presence of viral pathogens in natural areas a survey was conducted on an opportunistic sample of fifty eight wild (Felis silvestris silvestris) and feral cats (F. s. catus). The biological materials included serum, lung tissue extract and stool. Feline leukemia virus p27 antigen was detected in 13/50 serum/lung tissue extract samples (26%), canine distemper virus antibodies were detected in 2/26 serum/lung tissue extract samples (7.7%), feline coronavirus RNA was present in 6/29 stool samples (20.7%) and feline parvovirus DNA in 2/29 stool samples (6.9%). Canine distemper virus RNA was not detected. Feline immunodeficiency virus and feline coronavirus antibodies were not detected. Evidence of exposure to feline leukemia virus, canine distemper virus, feline coronavirus and feline parvovirus was found in wild and feral cats raising the importance of performing a comprehensive survey to correctly evaluate the potential threat of infectious diseases to endangered species, namely to the wildcat and to the Iberian lynx, which is meant to be reintroduced after 2012 in Portugal.

Prevalence of Feline Coronavirus Antibodies in Japanese Domestic Cats during the Past Decade

From 2001 to 2010, 17,392 Japanese cats were examined for feline coronavirus (FCoV) antibodies. The seroprevalence of purebreds (66.7%) was higher than that of random breds (31.2%). Seroprevalence increased greatly in purebreds by three months of age, while it did not fluctuate greatly in random breds with aging, indicating that cattery environments can contribute to FCoV epidemics. Purebreds from northern regions of Japan were likely to be seropositive (76.6% in Hokkaido, 80.0% in Tohoku), indicating cattery cats in cold climates might be more closely confined. Among purebreds, the American shorthair, Himalayan, Oriental, Persian, and Siamese showed low seroprevalence, while the American curl, Maine coon, Norwegian forest cat, ragdoll and Scottish fold showed high seroprevalence. There would also be breed-related differences in Japan similar to the previous studies in Australia.

Prevalence of feline coronavirus antibodies in cats in Bursa province, Turkey, by an enzyme-linked immunosorbent assay

Feline sera from Bursa province (Turkey) were assayed for coronavirus antibody using an enzyme-linked immunosorbent assay (ELISA). The study was performed on 100 sera collected from cats belonging to catteries or community shelters and to households. The serum samples were initially tested with the virus neutralisation (VN) test and the results were then compared with the ELISA. The VN yielded 79 negative and 21 positive sera but the ELISA confirmed only 74 as negative. The ELISA-negative sera were also found to be free of feline coronoviruses-specific antibodies by Western blotting. Using the VN as the gold standard test, ELISA had a sensitivity of 100% and a specificity of 93.6%, with an overall agreement of 95%. The Kappa (κ) test indicated high association between the two tests (κ = 0.86, 95% confidence interval (CI) 0.743–0.980). The positive predictive value (PPV) was 0.8, and the negative predictive value (NPV) was 0.93. The prevalence of FCoV II antibodies in the sampled population based on the gold standard was 62% (95% CI 0.44–0.77) among multi-cat environments, and 4% (95% CI 0.01–0.11) among single cat households.

Prevalence of Feline Coronavirus Antibodies in Cats in Bursa (Turkey) by An Enzyme-Linked Immunosorbent Assay

Since feline coronavirus (FCoVs) are ubiquitous pathogens, the seroprevalence was studied to monitor infection in Turkey. One hundred sera, collected from cats belonging to catteries or community shelters and to households, were tested. The VN test was performed and compared with ELISA. Each sample was heat-inactivated and serial two-fold dilutions starting from 1/2 were mixed with 100TCID50/50μl of FCoV type II strain 25/92. Subsequently, freshly trypsinized CRFK cells were added and the plates were incubated at 37°C for 96h.

Comparison of Serologic Techniques for the Detection of Antibodies against Feline Coronaviruses

The seroprevalence of feline coronavirus (FCoV) antibodies was studied in cats in southern Italy. One hundred twenty sera collected from cats belonging to catteries or community shelters and to households were tested for FCoV type I and II antibodies. The virus neutralization (VN) was performed and compared with indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA). Ninety-six sera tested positive for FCoV antibodies by VN and ELISA. Interestingly, ELISA revealed 2 more positive sera than did the VN test and 3 more positive sera than did the IFAT. All results were confirmed by Western blotting. ELISA proved to be more sensitive and detected a seroprevalence of about 82%. Considering the cross-reactivity of FCoV type I and type II, ELISA was able to detect antibodies against both serotypes, allowing the use of the assay as a reference test for sera screening. The high prevalence of antibodies observed indicates that FCoVs are common in southern Italian cat populations.

Seroprevalence study of feline coronavirus in owned and feral cats in Sydney, Australia.

Objectives To establish the seroprevalence of Feline Coronavirus (FCoV) infection in two defined groups of cats in Sydney: owned and feral cats; ii) to identify factors associated with an increased risk of infection with FCoV; and iii) to establish the seroprevalence and FCoV antibody titres of owned cats with immunohistochemically confirmed feline infectious peritonitis (FIP). Design Prospective multi‐institutional cross sectional study. Procedure Serum samples from owned cats presented to three inner city veterinary clinics in Sydney and feral cats from a colony in South Western Sydney over an 11‐month period were tested for FCoV antibodies using the Immunocomb test kit. The relationship between serological score and six major factors (breed, age, gender, number of cats per household, living environment and health status) in the owned cat sample population was analysed and compared to cats with FIP. Results The seroprevalence of FCoV infection in the sample population of owned and feral cats was 34% and 0%, respectively. The median Immunocomb scores of DSH, Persian, Siamese and Devon Rex cats were significantly lower than that of Burmese, BSH, Abyssinian, Birman, Ragdoll and Russian Blue. The median Immunocomb score of pedigree cats less than 2 years‐of‐age was significantly higher than for pedigree cats greater than 2 years‐of‐age. This distinction was not evident in DSH cats in these age groups. The number of cats per household at the time of blood collection had a strong positive association with Immunocomb score. The median Immunocomb score of cats with immunohistochemically confirmed FIP was significantly higher than cats in the sample population of owned cats but there was sufficient overlap between these two groups to make definitive diagnosis of FIP by serology impossible. Conclusion This represents the first seroprevalence study of FCoV in Australia. The major determinants of antibody score of owned cats identified in this study were breed, age and the number of cats per household. The significant relationship between the breed of the cat and the FCoV antibody titre further supports the notion, proposed previously by the authors, that breed related differences exist in the immunological response to FCoV infection.

The relationship between the Feline Coronavirus antibody titre and the age, breed, gender and health status of Australian cats

To investigate the relationship between Feline Coronavirus (FCoV) antibody titres and age, breed, gender and health status of Australian cats Retrospective study Results from two serological tests that measure FCoV antibody levels, the Coronase test and the 7B Feline Infectious Peritonitis (FIP) test, were recorded over a 2-year period, with patient signalment, history, presenting complaint and the reason for ordering the test (as available). Results from each antibody test were related to four explanatory variables (breed, age, gender and health status at the time of blood collection) using univariate ordinal logistic regression analyses, Mann Whitney U tests, one-sample sign tests or Kruskal-Wallis analyses, as appropriate. Results from 637 Coronase and 191 7B FIP antibody tests were recorded. There were significant differences in median Coronase antibody titres between breeds of cats (P < 0.0005). Specifically, the median Coronase antibody titres of Siamese, Persians, Domestic Shorthairs and Bengal cats (100) were significantly lower than that of British Shorthairs, Cornish Rex and Burmese cats (400, P < 0.0005). There was no statistical relationship between the Coronase or 7B FIP antibody titres and age, gender or overall health status, even when considering only those cats in which clinical signs suggestive of FIP were present. This study reinforces the complexity of interpreting serological tests for FCoV in both healthy cats and patients with signs compatible with FIR Unique to this study is the detection of a significant relationship between breed and median FCoV antibody titre. This supports the theory that breed related differences exist in response to FCoV infection. The distribution of median Coronase antibody titres by breed was very similar to the pattern of breed predisposition to FIP recently reported in Sydney.

Evaluation of an in-practice test for feline coronavirus antibodies

A commercially available in-practice test for feline coronavirus (FCoV) antibodies (FCoV Immunocomb, Biogal Galed Laboratories) was evaluated by comparison with the gold standard FCoV immunofluorescent antibody (IFA) test. One hundred and three serum or plasma samples were selected and tested: 70 were positive by both tests, 24 were negative by both tests. The in-practice test produced five false positive and four false negative results. The sensitivity of the in-practice test was 95% and the specificity was 83%. When the titres were compared it was found that the in-practice test results were significantly correlated with IFA titres but the degree of correlation was not likely to be clinically useful. The IFA titres of the four false negative samples were found to be low (less than 40) which suggests that even a cat with a false negative result is still unlikely to be excreting FCoV. A negative result with the in-practice assay is likely to be reliable for screening cats prior to entry into an FCoV-free cattery or stud. It would also be useful in the investigation of suspected FIP as most cats with this condition have high IFA titres of antibodies. A strong positive result would be useful in the diagnosis of FIP (in conjunction with other biochemical and cytological testing), but positive results would be of limited value in monitoring FCoV infection in healthy cats as the antibody titre could not be reliably compared with those obtained with IFA. All positive results obtained using the in-practice kit should be confirmed and titrated by IFA. The kit also appeared to work efficiently with ascites samples ( n=6) but too few samples were analysed to draw firm conclusions.

High viral loads despite absence of clinical and pathological findings in cats experimentally infected with feline coronavirus (FCoV) type I and in naturally FCoV-infected cats

Specified pathogen-free cats were naturally infected with FCoV or experimentally infected with FCoV type I. Seroconversion was determined and the course of infection was monitored by measuring the FCoV loads in faeces, whole blood, plasma and/or monocytes. Tissue samples collected at necropsy were examined for viral load and histopathological changes. Experimentally infected animals started shedding virus as soon as 2 days after infection. They generally displayed the highest viral loads in colon, ileum and mesenteric lymph nodes. Seroconversion occurred 3–4 weeks post infection. Naturally infected cats were positive for FCoV antibodies and monocyte-associated FCoV viraemia prior to death. At necropsy, most animals tested positive for viral shedding and FCoV RNA was found in spleen, mesenteric lymph nodes and bone marrow. Both experimentally and naturally infected cats remained clinically healthy. Pathological findings were restricted to generalized lymphatic hyperplasia. These findings demonstrate the presence of systemic FCoV infection with high viral loads in the absence of clinical and pathological signs.

Detection of feline coronavirus antibody, feline immunodeficiency virus antibody, and feline leukemia virus antigen in ascites from cats with effusive feline infectious peritonitis.

To investigate the usefulness of ascites as a material for viral tests in cats with effusive feline infectious peritonitis (FIP), we attempted to detect anti-feline coronavirus antibody, anti-feline immunodeficiency virus antibody, and feline leukemia virus antigen in ascites from 88 cats clinically suspected with effusive FIP. In each of these three viral tests, all cats positive for serum antibody/antigen were also positive for ascitic antibody/antigen, while cats negative for serum antibody/antigen were also negative for ascitic antibody/antigen. This finding indicates that ascites is useful for these viral tests.