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Guidelines for performing single-platform absolute CD4+ T-cell determinations with CD45 gating for persons infected
Francis F Mandy1, Janet K A Nicholson, J Steven McDougal
1Bureau of HIV/AIDS, STD and TB, Population and Public Health Branch, Health Canada, Ottawa, Ontario, Canada.
Insights
New CDC guidelines detail single-platform technology (SPT) for enumerating CD4+ T-cells and lymphocyte subsets in human immunodeficiency virus (HIV) infection. This method uses one instrument for accurate absolute counts, enhancing HIV monitoring.
Area of Science:
- Clinical laboratory science
- Immunology
- Virology
Background:
- Guidelines for CD4+ T-cell enumeration are crucial for monitoring human immunodeficiency virus (HIV) infection.
- Previous recommendations (1997) focused on dual-platform technology (DPT).
- Advancements necessitate updated guidelines for laboratory professionals.
Framework:
- Introduces single-platform technology (SPT) for lymphocyte subset enumeration.
- SPT utilizes a single instrument and a single tube for analysis.
- Employs internal calibrator beads and three- or four-color flow cytometry.
- Incorporates CD45 gating for calculating absolute counts and percentages.
Implementation:
- Provides practical guidance for laboratorians performing immunophenotyping.
- Addresses specific concerns related to SPT implementation.
- Covers essential laboratory safety and specimen handling protocols.
Implications:
- Facilitates accurate and efficient CD4+ T-cell count determination.
- Supports improved monitoring of HIV disease progression and treatment efficacy.
- Enhances laboratory workflow and potentially reduces costs compared to DPT.
Abstract:
These guidelines were developed by CDC for laboratorians who perform immunophenotyping for detection and enumeration of CD4+ T-cells and other lymphocyte subsets in persons infected with human immunodeficiency virus (HIV). The guidelines describe single-platform technology (SPT), a process in which absolute counts of lymphocyte subsets are measured from a single tube by a single instrument. SPT incorporates internal calibrator beads of known quantity in the analysis of specimens by three- or four-color flow cytometry. With CD45 gating, the relative numbers of beads and lymphocyte subsets are enumerated, and their absolute numbers and percentage values are calculated. This report supplements previous recommendations published in 1997 (CDC. 1997 revised guidelines for performing CD4+ T-cell determinations in persons infected with human immunodeficiency virus [HIV]. MMWR 1997;46[No. RR-2]) that describe dual-platform technology, a method in which absolute counts are derived from measurements obtained from two instruments--a flow cytometer and hematology analyzer. The new recommendations address concerns specific to the implementation of SPT as well as other general topics such as laboratory safety and specimen handling.
