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BACKGROUND: Rapid expansion of antiretroviral therapy in Malawi has occurred in the relative absence of suitable pediatric CD4 counting facilities. We have recently validated in adults a simplified affordable flow cytometric CD4 counting method, the Blantyre count. There is a need for this technology to transfer to government laboratories run by local staff, and to be validated in children, where %CD4/lymphocyte values are required. METHODS: We assessed agreement of %CD4/lymphocyte values determined by the Blantyre count and Panleucogate methods on an EPICS XL-MCL flow cytometer on 113 venous blood samples from HIV-seropositive children in Blantyre, Malawi. All assays were performed by two Malawian laboratory technicians. RESULTS: Overall bias between the two methods was -0.13% (95% CI -0.37 to 0.11) and limits of agreement were -2.69 to 2.43% (95% CI -3.11 to -2.27 and 2.01 to 2.85). Limits of agreement were within -3.00 and 3.00 for each laboratory technician. Coefficient of variation for the Blantyre count assay was 2.0% and samples showed good stability over 5 days. CONCLUSIONS: The Blantyre count method can accurately determine %CD4/lymphocyte values in blood of HIV-seropositive children on an EPIC XL-MCL flow cytometer at a reagent cost of US $0.21 per test or less. The assay can be competently carried out by local laboratory technicians.

Original publication

DOI

10.1002/cyto.b.20411

Type

Journal article

Journal

Cytometry B Clin Cytom

Publication Date

2008

Volume

74 Suppl 1

Pages

S90 - S97

Keywords

Bias, Blood Specimen Collection, CD4 Lymphocyte Count, Child, Child, Preschool, Female, Flow Cytometry, Humans, Infant, Malawi, Male, Observer Variation, Reproducibility of Results, Software