Simplified Volumetric Flow Cytometry Allows Feasible and Accurate Determination of Cd4 T Lymphocytes in Immunodeficient Patients Worldwide

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The determination of CD4 cells is of crucial clinical importance for patients with AIDS. However, the high costs involved represent limitations for CD4 cell counting in developing countries. In order to provide an affordable technique, we introduced a simplified volumetric counting (SVC) technique without sample manipulations and investigated it in a multicentre study. Blood samples from 434 healthy donors and immunodeficient patients were tested in eight hospital laboratories in Europe, Africa and Asia. CD4 cell counts were compared using in-house flow cytometric methods and the SVC technique. The SVC method was performed on a low-cost flow cytometer (CyFlow SL, Partec, Münster, Germany) after 15 min antibody incubation without pre-analytic manipulations, such as washing or erythrocyte lysing procedures. Linear regression analysis demonstrated a correlation of r=0.942 (Europe), r=0.952 (Africa) and r=0.989 (Asia) between the SVC technique and the in-house methods. Bland Altman plot analysis of all patient data showed a mean bias between the two methods of +26 CD4 cells in favour of the SVC technique (measured range: 6-1905 cells/microl; median CD4 cell count: 388/microl). Three centres used the FACS-count technique (Becton-Dickinson, San José, Calif., USA) as an in-house method dispensing with pre-analytic manipulations. The comparison of SVC and FACS-count method revealed a mean bias of +32 CD4 cells/microl (median CD4 cell count: 349/microl). The accuracy of the SVC was tested on standards with known CD4 cell counts (n=6) and was shown to be 95.2%. The low-cost device and the simplified no-lyse, no-wash test procedure reduces the costs per determination and facilitates the use of flow cytometry in developing countries.

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