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Immunoassay for detection of feline immunodeficiency virus core antigen
G K Tilton1, T P O'Connor, C L Seymour
1IDEXX Corporation, Portland, Maine 04101.
Insights
A new enzyme-linked immunosorbent assay detects feline immunodeficiency virus (FIV) p24 antigen. This sensitive FIV test confirms the virus in infected cat cells.
Area of Science:
- Veterinary Virology
- Immunology
- Molecular Diagnostics
Background:
- Feline immunodeficiency virus (FIV) is a significant lentivirus affecting domestic cats globally.
- Accurate diagnostic methods are crucial for managing FIV in feline populations.
Purpose of the Study:
- To develop a sensitive enzyme-linked immunosorbent assay (ELISA) for detecting FIV group-associated antigen (gag) p24.
- To establish a reliable method for confirming FIV presence in cell cultures.
Main Methods:
- A monoclonal antibody-based ELISA in a microdilution plate format was designed.
- Sensitivity was determined using serially diluted disrupted FIV samples.
- A qualitative antibody inhibition assay was coupled for confirmation.
Main Results:
- The assay demonstrated a sensitivity limit of approximately 0.2 ng/ml for FIV p24.
- The FIV p24 ELISA was approximately eightfold more sensitive than reverse transcriptase assays.
- The test successfully detected and confirmed FIV in cultured feline lymphocytes.
Conclusions:
- The developed ELISA is a highly sensitive and specific tool for FIV p24 detection.
- This assay provides a reliable method for diagnosing FIV in clinical and research settings.
- The coupled confirmation assay enhances diagnostic accuracy for feline lentivirus infections.
Abstract:
The feline immunodeficiency virus (FIV) is a recently identified feline lentivirus that has been found at significant levels in domestic cat populations worldwide. A microdilution plate format, monoclonal antibody-based enzyme-linked immunosorbent assay was developed for the detection of the FIV group-associated antigen (gag) designated p24. Assays of serially diluted samples containing disrupted virus showed that the assay had a sensitivity limit of approximately 0.2 ng/ml for FIV p24. The assay was approximately eightfold more sensitive than the assay for viral reverse transcriptase activity when it was tested with diluted tissue culture samples. A qualitative confirmation assay by standard antibody inhibition techniques was coupled to the screening test methodology. The test was used to detect and confirm the presence of virus in cultured feline lymphocytes from infected animals.
