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CD4 lymphocyte enumeration in patients with human immunodeficiency virus infection using three-colour and four-colour
1Department of Haematology-Oncology, National University Hospital, 5 Lower Kent Ridge Road, Singapore 119074. LauLeeGong@nuh.com.sg
Insights
CD4 lymphocyte percentages showed good agreement between two flow cytometry methods. However, absolute CD4 counts did not agree well, limiting clinical interchangeability between hospitals.
Area of Science:
- Clinical immunology
- Flow cytometry
- HIV diagnostics
Background:
- Accurate CD4 lymphocyte enumeration is critical for managing HIV infection.
- Dual-platform flow cytometry is widely used but requires validation between laboratories.
Purpose of the Study:
- To evaluate the correlation and agreement of CD4 lymphocyte enumeration methodologies between two major hospitals.
- To assess the comparability of dual-platform flow cytometry results in clinical practice.
Main Methods:
- Analysis of 52 blood samples from HIV-infected patients using dual-platform flow cytometry.
- Comparison of 3-colour (Hospital A) and 4-colour (Hospital B) flow cytometry techniques.
- Statistical analysis using linear regression and Bland-Altman plots.
Main Results:
- High correlation (r > 0.95) was observed for total white cell counts, absolute lymphocyte counts, and CD4 lymphocyte percentages.
- CD4 lymphocyte percentages showed good agreement (mean difference 0.05%).
- Absolute CD4 lymphocyte counts demonstrated poor agreement (mean difference 14/uL, wide limits of agreement).
Conclusions:
- CD4 lymphocyte percentages obtained by both methods are comparable for clinical use.
- Absolute CD4 lymphocyte counts from the two hospitals are not interchangeable due to poor agreement.
- Inter-laboratory comparability of absolute CD4 counts is a concern for clinical decision-making in HIV management.
Introduction:
This study aims to evaluate the correlation and agreement between 2 methodologies of CD4 lymphocyte enumeration.
Materials And Methods:
Fifty-two blood samples from patients with human immunodeficiency virus (HIV) infection were sent for CD4 lymphocyte enumeration at 2 major hospitals using dual-platform flow cytometry where the absolute lymphocyte counts were determined on separate haematology analyzers. CD4 cell enumeration was accomplished using 3-colour flow cytometry on the FACSCalibur cytometer (Becton Dickinson) in Hospital A and 4-colour flow cytometry on the Coulter Epic XL cytometer (Beckman Coulter) in Hospital B. The percentages and absolute counts of CD4 lymphocytes obtained were analysed using both linear regression and Bland-Altman plots.
Results:
On linear regression plots, the total white cell counts, absolute lymphocyte counts, CD4 lymphocyte percentages and absolute CD4 counts from the 2 hospitals correlated well with correlation coefficients, r > 0.95. On the Bland-Altman plots, there was a mean difference of 0.13 x 10(9)/L and 0.06 x 10(9)/L in the total white cell and absolute lymphocyte counts from the 2 hospitals respectively. The CD4 lymphocyte percentages revealed a mean difference of only 0.05% (95% limits of agreement, -3.6 to 3.7%) but there was a mean difference of 14/uL in the absolute CD4 lymphocyte counts (95% limits of agreement, -113 to 142/uL).
Conclusions:
The CD4 lymphocyte percentages obtained using the 3-colour and 4-colour flow cytometry correlated and agreed well. However, the absolute CD4 lymphocyte counts obtained using the dual-platform technique in both the hospitals did not agree well. Hence, absolute CD4 lymphocyte counts from the 2 hospitals cannot be used interchangeably in clinical practice due to poor inter-laboratory comparability.
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