Comparative study of diagnostic testing for feline leukemia virus infection.

Results of commercially available diagnostic test kits and commercial laboratory test results were compared for ability to detect FeLV antigen. Results of the immunofluorescent antibody (IFA) test were compared with test kit ELISA results and with results of a system in which samples were applied to an absorbent material, dried, sent to a laboratory, eluted, and assayed by a plate ELISA. Test kits were generally highly sensitive and specific, compared with the IFA test performed at a commercial laboratory. Feline heterophile antibody, specific for mouse immunoglobulin, was detected in approximately 0.14 to 0.57% of the cat population. Test kits B, E, and D contain reagents that correct for antimouse antibodies. During 1989 and 1990, 2,229 feline serum samples were tested for FeLV antigen (gsa p27); positive ELISA results were obtained for 204 (9%) of the samples. Results for 32 (1.4%) samples were interpreted as equivocal (color development slightly exceeded that of the negative control, but was much less than that of the positive control). Collectively, the data indicate that when testing serum or saliva, a negative test result may be a good predictor that a cat is not infected. In populations of cats in which FeLV prevalence is low, a positive test result may not be reliable and thus, a confirmatory test should be performed.     

Prevalence and sequelae of feline leukemia virus infection in laboratory cats

Over a 4-year period, 1,683 pound-source cats received at a research institution were screened for feline leukemia virus (FeLV) infection, using an indirect fluorescent antibody test. Viremia was detected in 83 of the cats, for a prevalence of 4.9%. During this period, FeLV infection was detected in 5 kittens on a research project; lymphoma or anemia developed 6 to 17 months after the infections were detected. It was concluded that apparently healthy cats infected with FeLV may not be appropriate for some biomedical research projects.     

Long-term impact on a closed household of pet cats of natural infection with feline coronavirus, feline leukaemia virus and feline immunodeficiency virus

A closed household of 26 cats in which feline coronavirus (FCoV), feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV) were endemic was observed for 10 years. Each cat was seropositive for FCoV on at least one occasion and the infection was maintained by reinfection. After 10 years, three of six surviving cats were still seropositive. Only one cat, which was also infected with FIV, developed feline infectious peritonitis (FIP). Rising anti-FCoV antibody titres did not indicate that the cat would develop FIP. The FeLV infection was self-limiting because all seven of the initially viraemic cats died within five years and the remainder were immune. However, FeLV had the greatest impact on mortality. Nine cats were initially FIV-positive and six more cats became infected during the course of the study, without evidence of having been bitten. The FIV infection did not adversely affect the cats' life expectancy.     

Demodicosis in two cats seropositive for feline immunodeficiency virus

Two cats were diagnosed with generalized demodicosis. Serotest results were negative for FeLV and positive for feline immunodeficiency virus. In one cat, demodicosis resolved in response to topical application of lime-sulfur solution, but the other cat was euthanatized.     

Efficacy of a Feline Leukemia Virus Vaccine in a Natural Exposure Challenge

A commercial feline leukemia virus (FeLV) vaccine was evaluated in a natural exposure system. All kittens were negative for FeLV antigen on two enzyme-linked immunosorbent assay (ELISA) tests and one indirect immunofluorescence antibody (IFA) test before vaccination or exposure. Twenty-three kittens were vaccinated subcutaneously at nine and 12 weeks of age. The vaccinated kittens and 14 unvaccinated littermates were housed in an infected environment starting at 14 weeks. The kittens were exposed for 24 weeks by living in a large room with one feline leukemia virus-positive, asymptomatic adult cat for each five kittens. Sixty-four percent of the unvaccinated kittens and 70% of the vaccinated kittens became infected as determined by ELISA. Forty-three percent of unvaccinated kittens and 39% of vaccinated kittens died. There was no difference between the infection and mortality of vaccinated kittens that developed antibodies to anti-FeLV glycoprotein 70-envelope antigen and those that did not. Consideration should be given to evaluation of feline leukemia virus vaccines using "street" virus in a natural exposure system.     

Ten-year study comparing enzyme-linked immunosorbent assay with the immunofluorescent antibody test for detection of feline leukemia virus infection in cats.

Immunodetection tests for feline retroviruses are powerful tools used in modern veterinary practice. Veterinarians must fully understand the characteristics--strengths and weaknesses--of the FeLV tests so that the information gained from them can be used properly. Any FeLV ELISA or immunofluorescent antibody (IFA) test is a method for detection of FeLV infection (the virus) and is not a diagnostic test for leukemia or other feline disease. From previous studies, it was determined that the most accurate test for detection of persistent FeLV infection is the IFA test, which detects FeLV antigens in cytoplasm of leukocytes in the blood of infected cats. In the study reported here, 1,142,600 FeLV IFA tests were performed between June 1972 and December 1990. During this period 19.8% of the IFA test results were positive and 78% were negative. Evaluation was not possible for the remaining 2.2% of the tests because of lack of enough leukocytes in the smears to evaluate, or nonspecific staining reactions. In 1979, 7 years after introduction of the IFA test, in-hospital FeLV ELISA were introduced, which enabled veterinarians to test for FeLV in their hospitals. Ever since that time, continual discrepancies have been reported between results of FeLV ELISA and IFA tests, particularly between positive ELISA results and their IFA test confirmation. A 10-year comparison was made between practitioner-performed in-hospital FeLV ELISA (n = 20, 240 tests) results and FeLV IFA test performed by a commercial laboratory. All samples tested by ELISA were submitted (for confirmation of results) by veterinarians from the United States, Canada, Europe, Japan, and Australia.(ABSTRACT TRUNCATED AT 250 WORDS)     

The presence of leukaemia (lymphosarcoma) and feline leukaemia virus (FeLV) in cats in The Netherlands

To study the presence and spread of feline leukaemia virus (FeLV) in The Netherlands, seven different groups of cats were examined. The indirect fluorescent antibody (IFA) test was used to detect FeLV-antigen in blood smears. Of cats with lymphosarcoma/leukaemia 73.2% were positive and 32.4% with infectious peritonitis were positive. Only one of sixty-six cats with other tumours–a cat with mammary carcinoma–was positive.
Forty-two (7.5%) of 557 cats with various complaints were positive for FeLV-antigen. The IFA-test appeared to be an important diagnostic supplement.
Of all stud males which had had contact with FeLV-positive cats 24.7% were positive for FeLV-antigen, whereas all stud males which had not had this contact, were negative.
There was a distinct difference between the percentages of FeLV-positive individuals in the groups of cats which had had (20.6%) and which had not had (0.4%) contact with FeLV-positive cats.
From the follow-up study it was found that 67.3% of the FeLV-positive cats died from, or were destroyed because of, FeLV-associated diseases within a period of 20 months.     

Tumor necrosis factor alpha levels in cats experimentally infected with feline immunodeficiency virus: effects of immunization and feline leukemia virus infection.

Tumor necrosis factor alpha (TNF alpha) levels were determined by enzyme-linked immunosorbent assay (ELISA) and by cell culture bioassay in supernatants of lipopolysaccharide-stimulated feline monocyte cultures and in cat serum samples. There was a good correlation between the results obtained by the two methods. From the fact that TNF alpha was neutralized quantitatively by antibodies to human TNF alpha in feline monocyte supernatants and in feline sera, it was concluded that feline TNF alpha immunologically cross-reacts with human TNF alpha and that the human TNF alpha ELISA can be used to quantitate feline TNF alpha. During the first 6 months after experimental feline immunodeficiency virus (FIV) infection no differences in serum TNF alpha values were observed between infected and non-infected cats. TNF alpha levels increased significantly after primary vaccination with a feline leukemia virus (FeLV) vaccine in FIV infected cats over those in the non-infected controls. During secondary immune response TNF alpha levels rose transiently for a period of a few days in both the FIV positive and the FIV negative cats. After FeLV challenge, TNF alpha levels increased in all animals challenged with virulent FeLV for a period of 3 weeks. This period corresponded to the time necessary to develop persistent FeLV viremia in the control cats. It was concluded from these experiments that in the asymptomatic phase of FIV infection no increased levels of TNF alpha are present, similar to the situation in asymptomatic HIV infected humans. Activation of monocytes/macrophages in FIV infected cats by stimuli such as vaccination or FeLV challenge readily leads to increased levels of TNF alpha.     

A micro-enzyme-linked immunosorbent assay (ELISA) test for detection of feline leukemia virus in cats

The performance of a micro ELISA test for detection of feline leukemia virus (FeLV) infection was evaluated. The test was found specific for FeLV and feline sarcoma virus (FeSV) group-specific antigens in blood, plasma or serum of infected cats. Other common feline pathogens were negative to the test.Quantities as little as 7.8 ng of p-27 (the major group specific antigen of FeLV) per ml of sample gave positive results. The correlation between the micro ELISA test and the indirect immunofluorescent test commonly used for diagnosis of FeLV infection was 98% in 116 clinical cases and 184 samples from cats inoculated with FeLV and 100% in 100 specific pathogen-free cats.     

Lymphosarcoma in experimentally induced feline immunodeficiency virus infection [corrected]

A cat experimentally infected with feline immunodeficiency virus (FIV) but known to be free of feline leukaemia virus (FeLV) developed lymphosarcoma. The lesions in the liver and kidneys were present nine months after infection, when the cat was 21 months old. The cat had no overt signs of immunodeficiency and it is suggested that the B cell activation induced shortly after FIV infection produced a large pool of proliferating lymphocytes from which the malignant cells emerged.     

La leucose feline: Developpements recents sur l''etiologie et l''immunoprevention de la maladie

Feline leukemia is a useful model for malignant hematopoïetic tumor studies. It is caused by a type C, RNA virus, the Feline Leukemia virus (FeLV), transmitted horizontally, and widespread in the cat population.The presence of DNA sequences and virus specific RNA expression in cell cultures of SPF cats and cat embryos, indicates a vertical transmission may occur.These FeLV-related sequences in virus negative lymphosarcoma, almost from older cats, indicate that in certain FeLV related diseases the viral replication may not occur. An endogenous ecotropic feline virus may also explain this finding. The absence of FeLV gene expression in some lymphomatous cats—many older—suggest that, in these cats, spontaneous lymphoma may not be caused by FeLV.The widespread occurrence of feline xenotropic endogenous virus RD-114 gene, in feline lymphoma, suggest that expression of certain functions of this virus may be involved etiologically in the development of lymphoid tumors in the cat.Nevertheless, immunisation against FeLV would provide a good prevention against the main part of the feline lymphosarcomas and other FeLV-related diseases. Inactivated FeLV does not provide a good immunisation in young cats. By contrast a good protection against tumoral development is obtained by vaccination using the Feline oncogenic virus cell membrane antigen (FOCMA).     

Two cases of FeLV-associated dermatoses

Two cases of feline leukaemia virus (FeLV)-associated dermatosis are described. The first cat was affected by an ulcerative dermatitis identified as a giant-cell dermatosis. The second case was a cutaneous lymphoma. In both cases, FeLV antigens and FeLV genome were demonstrated in the affected skin immunologically and with polymerase chain reaction, respectively. The first case suggests that, like other retroviruses, at least some strains of FeLV can induce syncytium formation. As FeLV antigens and genome were demonstrated in a serologically negative cat, the second case suggests that focal skin FeLV replication may occur. FeLV-associated dermatoses are rare skin conditions that may be under-diagnosed.     

Prevalence of canine distemper virus, feline immunodeficiency virus and feline leukemia virus in captive African lions (Panthera leo) in Japan

Sero-prevalences of canine distemper virus (CDV), feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) were evaluated in 20 captive lions in two Japanese zoos. Anti-CDV antibody was detected in 13 of 20 lions. We could pursue antibody responses against CDV in three lions back to 1996. Sera collected in 1996 were negative for anti-CDV antibody, therefore, all of them showed sero-conversion in 2000. This result suggested that the epidemic of CDV infection in this zoo might have happened between 1996 and 2000. The lions were also examined for FIV and FeLV infections. We had no evidence for FeLV infection but eight lions were sero-positive for anti-FIV antibody.     

Exposure of cats to low doses of FeLV: seroconversion as the sole parameter of infection

In felids, feline leukemia virus (FeLV) infection results in a variety of outcomes that range from abortive (virus readily eliminated and never detectable) to progressive infection (persistent viremia and viral shedding). Recently, a novel outcome was postulated for low FeLV infectious doses. Naïve cats exposed to faeces of persistently infected cats seroconverted, indicating infection, but remained negative for provirus and p27 antigen in blood. FeLV provirus was found in some tissues but not in the bone marrow, infection of which is usually considered a necessary stage for disease progression. To investigate the impact of low FeLV doses on young cats and to test the hypothesis that low dose exposure may lead to an unknown pathogenesis of infection without involvement of the bone marrow, 21 cats were infected oronasally with variable viral doses. Blood p27, proviral and viral loads were followed until week 20 post-infection. Tissue proviral loads were determined as well. The immune response was monitored by measuring FeLV whole virus and p45 antibodies; and feline oncornavirus-associated cell membrane antigen (FOCMA) assay. One cat showed regressive infection (transient antigenemia, persistent provirus-positivity, and seroconversion) with provirus only found in some organs at sacrifice. In 7 of the 20 remaining cats FOCMA assay positivity was the only sign of infection, while all other tests were negative. Overall, the results show that FeLV low dose exposure can result in seroconversion during a presumed abortive infection. Therefore, commonly used detection methods do not detect all FeLV-infected animals, possibly leading to an underestimation of the prevalence of infection.     

Year two of follow-up evaluation of a randomized, blind field trial of a commercial feline leukemia virus vaccine.

A blind randomized field trial of a commercial FeLV vaccine was conducted. Cats on study were vaccinated with either a commercial FeLV vaccine or a placebo, then housed with FeLV-positive cats in a ratio of approximately 2 study cats to 1 infected cat (results of the first 12 months of the study have been reported). All surviving placebo-treated and FeLV-vaccinated cats were re-vaccinated 1 year after initial exposure to FeLV-infected cats. Exposure continued for an additional 12 months, and the viremia status of the cats was monitored by immunofluorescent antibody (IFA) and ELISA testing at 4-month intervals. During the second year of observation, 1 additional FeLV-vaccinated cat had positive results of 2 consecutive ELISA tests, but remained IFA negative. Classifying this cat as persistently viremic reduced the estimate of the preventable fraction, but did not alter the conclusions drawn earlier, viz, that vaccination appreciably reduces the number of cats that become persistently viremic after long-term natural exposure.     

Detection of feline leukaemia virus in blood and bone marrow of cats with varying suspicion of latent infection

The purpose of this study was to determine if polymerase chain reaction (PCR) could be used to detect FeLV proviral DNA in bone marrow samples of cats with varying suspicion of latent infection. Blood and bone marrow samples from 50 cats and bone marrow from one fetus were collected, including 16 cats with diseases suspected to be FeLV-associated. Serum enzyme-linked immunosorbent assay (ELISA), blood and bone marrow immunofluorescent antibody test (IFA), and blood and bone marrow PCR were performed on each cat, and IFA and PCR on bone marrow of the fetus. Forty-one cats were FeLV negative. Five cats and one fetus were persistently infected with FeLV. Four cats had discordant test results. No cats were positive on bone marrow PCR only. It appears persistent or latent FeLV infection is not always present in conditions classically associated with FeLV.     

Association of feline leukemia virus with lymphosarcoma and other disorders in the cat.

Two hundred fifty Boston cats with disorders such as lymphosarcoma, myeloproliferative disease, anemia, glomerulonephritis, pregnancy abnormalities, feline infectious peritonitis, toxoplasmosis, and various bacterial infections were examined for feline leukemia virus (FeLV) by immunofluorescence. Antibody titers against feline oncornavirus-associated cell membrane antigen (FOCMA) were tested in 133 of these cats. The tests for FeLV and FOCMA antibody were also conducted among healthy cats not known to have been exposed to FeLV, as well as among healthy cats from households where FeLV was known to be present. Most of the cats with lymphosarcoma and the other aforementioned disorders were infected with FeLV and low FOCMA antibody titers. Healthy cats known to have been exposed to FeLV were often viremic, but those that remained healthy were able to develop high FOCMA antibody titers. Healthy cats without known prior exposure to FeLV were unlikely to be viremic but often had detectable FOCMA antibody titers, indicating that some exposure occurs under natural conditions in the Boston area. The association of FeLV with infections other than lymphosarcoma was assumed to be caused by the immunosuppresive effect of FeLV, thus allowing development of disease.     

Serosurvey for feline leukemia virus and lentiviruses in captive small neotropic felids in S?o Paulo state, Brazil.

Feline leukemia virus (FeLV), Gammaretrovirus, and feline immunodeficiency virus, a Lentivirus, are members of the family Retroviridae, and may establish persistent infections in the domestic cat (Felis catus). Cytoproliferative and cytosuppressive disorders may result from infection with these viruses. Morbidity and mortality rates are high in domestic cats worldwide. Infection of endangered neotropic small felids with these viruses could be devastating. To investigate the prevalence of FeLV and feline lentiviruses in neotropic small felids kept in captivity in S?o Paulo state. Brazil, serum samples from 104 animals belonging to the species Leopardus pardalis, Leopardus tigrinus, Leopardus wiedii, Herpailurus yaguarondi, and Oncifelis geoffroyi were tested for FeLV and feline lentiviruses by commercially available immunoassays. All results were negative, suggesting that retrovirus infection is not an important clinical problem in these populations. Because domestic cats in S?o Paulo city are naturally infected with these pathogens, and feral cats are commonly found in zoologic facilities in Brazil, preventive measures should be taken to avoid transmission of retroviruses to naive populations of wild and captive neotropic felids in Brazil.     

Use of tears for diagnosis of feline leukemia virus infection

A comparison was made of the use of serum, tears, and saliva for the detection of feline leukemia virus (FeLV) infection in cats. Cotton swabs were used to collect saliva, and tear-test strips were used to collect tears. Specimens were analyzed by a commercially available ELISA. Using a 10- to 15-minute specimen incubation period, FeLV was detected in 70% of the saliva specimens and in 73% of the tear specimens from viremic (serum-positive) cats. Feline leukemia virus antigen was not detected in saliva and tear specimens from serum-negative cats. The sensitivity of the tear assay was improved by increasing the incubation time to 24 hours. Tear strips could be air-dried and stored at room temperature for up to 7 days without any appreciable loss of activity. Client-owned and experimentally infected laboratory cats were tested for FeLV, using air-dried tear-test strips and a 24-hour incubation period. Tears were positive (contained FeLV antigen) in 65 of 72 (90%) serum-positive cats and did not contain antigen in 46 of 46 (100%) serum-negative cats. Results of ELISA obtained from serum and tears also were compared with results obtained from indirect fluorescent antibody testing of blood smears. Results of indirect fluorescent antibody and ELISA compared favorably with each other and with the results of tear testing.