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Affordable CD4(+)-T-cell counting by flow cytometry: CD45 gating for volumetric analysis
George Janossy1, Ilesh V Jani, Nicholas J Bradley
1HIV Immunology, Department of Immunology and Molecular Pathology, Royal Free and University College Medical School, London, United Kingdom. janossy@rfhsm.u-net.com
Clinical and Diagnostic Laboratory Immunology
|September 3, 2002
Summary
This study introduces a cost-effective volumetric flow cytometry method for monitoring human immunodeficiency virus (HIV) disease. The new approach provides accurate CD4(+) T-cell counts, making it suitable for resource-poor settings.
Area of Science:
- Immunology
- Biomedical Engineering
- Public Health
Background:
- Accurate CD4(+) T-cell counts are crucial for monitoring human immunodeficiency virus (HIV) disease progression.
- Current industry-standard flow cytometers are often unaffordable for routine use in resource-poor settings.
- There is a need for cost-efficient and accessible methods for CD4(+) T-cell enumeration.
Purpose of the Study:
- To develop and evaluate a cost-efficient volumetric flow cytometry method for CD4(+) T-cell counting.
- To assess the agreement of the new method with existing industry standards.
- To determine the suitability of this method for large-scale use in resource-limited laboratories.
Main Methods:
- Combined volumetric flow cytometry with simpler protocols and generic monoclonal antibodies (MAbs).
- Generated absolute counts using CD45/CD4 and CD45/CD8 MAb combinations.
- Utilized Bland-Altman analyses to compare results with volumetric and bead-based systems.
Main Results:
- The volumetric method showed good agreement with industry standards for CD4(+) T-cell counts.
- Limits of agreement for CD4 counts were -29 to +46 cells/mm(3) with the CytoronAbsolute system.
- The method demonstrated high efficiency, with a potential throughput of over 30,000 samples per year.
Conclusions:
- Volumetric flow cytometry using generic CD45, CD4, and CD8 MAbs is a cost-efficient and accurate method for CD4(+) T-cell monitoring.
- This approach is highly suitable for large regional laboratories in resource-poor settings.
- The developed method offers a viable alternative for HIV disease management in underserved areas.