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p24 antigen capture assay for quantification of human immunodeficiency virus using readily available inexpensive
1Laboratory of Persistent, Viral Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, NIH, Hamilton, Montana 59840-2932, USA.
Insights
We developed a cost-effective antigen capture ELISA to quantify HIV-1 p24 protein, crucial for studying virus replication. This sensitive assay is also effective for detecting related viral proteins in HIV-2 and SIV.
Area of Science:
- Virology
- Immunology
- Biochemistry
Background:
- Antigen capture enzyme-linked immunosorbent assay (ELISA) is the standard for quantifying human immunodeficiency virus type-1 (HIV-1) p24 gag protein, indicating virus replication.
- Existing methods can be costly, limiting widespread research application.
Purpose of the Study:
- To describe a novel, inexpensive antigen capture ELISA for quantifying HIV-1 p24 gag protein.
- To provide optimization strategies for specific research needs.
- To assess the assay's sensitivity, linear range, and applicability to related viruses.
Main Methods:
- Development of an antigen capture ELISA using affordable reagents.
- Optimization of assay variables for enhanced performance.
- Validation of sensitivity and linear dose response.
Main Results:
- The developed ELISA demonstrates sensitivity comparable to commercial kits, with a limit of 25 pg/ml.
- The assay exhibits a linear dose response for p24 concentrations ranging from 25-1000 pg/ml.
- The assay effectively detects gag proteins from HIV-1, HIV-2, and simian immunodeficiency virus (SIV).
Conclusions:
- This cost-effective ELISA provides a sensitive and reliable method for quantifying HIV-1 p24 gag protein.
- The assay's affordability and broad applicability make it valuable for diverse research settings.
- The method is also suitable for detecting gag proteins of related immunodeficiency viruses.
Abstract:
Antigen capture enzyme-linked immunosorbent assay (ELISA) for quantitation of the p24 gag protein of human immunodeficiency virus type-1 (HIV-1) is currently the most common method used to demonstrate virus replication both in vivo and in vitro. The present paper describes an ELISA employing readily available inexpensive reagents and gives detailed suggestions for optimizing the variables for specific purposes. The assay is as sensitive as commercial kits (25 pg/ml) and has a linear dose response over a wide range of p24 concentrations (25-1000 pg/ml). For these reasons, as well as its low cost, this assay has proven useful in a variety of research applications. This assay also has been found to be effective in detecting the gag protein of human immunodeficiency virus type-2 and simian immunodeficiency virus.
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