J Vet Sci.  2014 Mar;15(1):91-97. 10.4142/jvs.2014.15.1.91.

Development and clinical evaluation of a rapid diagnostic kit for feline leukemia virus infection

• Kim WS
• Chong CK
• Kim HY
• Lee GC
• Jeong W
• An DJ
• Jeoung HY
• Lee JI
• Lee YK

• KMID: 1737614
• DOI: http://doi.org/10.4142/jvs.2014.15.1.91

Abstract

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 x 10(9) and 0.86 x 10(9), 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 x 10(4) 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.

Keyword

anti-FeLV p27 monoclonal antibody; feline leukemia virus; polymerase chain reaction; rapid diagnostic test; recombinant FeLV p27

MeSH Terms

Animals
Antibodies, Monoclonal/blood
Cats
Diagnostic Tests, Routine/*veterinary
Female
Gene Products, gag/*blood
Leukemia Virus, Feline/immunology/*isolation & purification
Leukemia, Feline/*diagnosis
Mice, Inbred BALB C
Sensitivity and Specificity
Antibodies, Monoclonal
Gene Products, gag

Figure

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  Fig. 1 (A) Cloning of the FeLV p27 gene. (B) Expression and purification of soluble p27 protein. M, (A) DNA size markers; (B) protein size markers; Lane 1, insoluble fraction from the cell lysate; Lane 2, soluble fraction from the cell lysate; Lane 3, purified recombinant p27 (rec. p27). S: sample.

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  Fig. 2 Antigenic activity test for the rec. p27 protein using SNAP FeLV Ag/FIV Ab.

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  Fig. 3 Reactivity of each hybridoma to rec. p27 protein (■) and purified FeLV (□).

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  Fig. 4 Analytical sensitivity of the rapid diagnostic test (RDT) using rec. p27 protein (A) and FeLV particles (B). IU/mL: infectious units per mL, +W: weakly positive.

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