Feline leukemia virus (FeLV) causes a range of neoplastic
and degenerative diseases in cats. To obtain a more sensitive
and convenient diagnosis of the disease, we prepared
monoclonal antibodies specific for the FeLV p27 to develop a
rapid diagnostic test with enhanced sensitivity and
specificity. Among these antibodies, we identified two clones
(hybridomas 8F8B5 and 8G7D1) that specifically bound to
FeLV and were very suitable for a diagnostic kit. The affinity
constants for 8F8B5 and 8G7D1 were 0.35 × 109 and 0.86 ×
109, respectively. To investigate the diagnostic abilities of the
rapid kit using these antibodies, we performed several
clinical studies. Assessment of analytical sensitivity revealed
that the detection threshold of the rapid diagnostic test was 2
ng/mL for recombinant p27 and 12.5 × 104 IU/mL for FeLV.
When evaluating 252 cat sera samples, the kit was found to
have a kappa value of 0.88 compared to polymerase chain
reaction (PCR), indicating a significant correlation between
data from the rapid diagnostic test and PCR. Sensitivity and
specificity of the kit were 95.2% (20/21) and 98.5% (257/261),
respectively. Our results demonstrated that the rapid
diagnostic test would be a suitable diagnostic tool for the
rapid detection of FeLV infection in cats.