Low-CD4+ T-lymphocyte blood samples for external quality assessment of CD4 testing in resource-poor settings

Abstract

© 2014, Springer-Verlag Berlin Heidelberg. External quality assessment (EQA) of CD4 testing is important for clinically monitoring of patients with HIV/AIDS. EQA materials are limited to blood samples from normal blood donors with normal CD4+ T-lymphocyte levels. This study aimed to develop low-CD4+ T-lymphocyte blood samples for CD4 EQA. CD4+ T-lymphocyte-depleted blood samples were prepared using a magnetic bead separation technique. These CD4+ T-lymphocyte-depleted blood samples were mixed with undepleted whole blood samples to obtain low-CD4+ T-lymphocyte blood samples. The percentage and the number of CD4+ T-lymphocytes were determined using a flow cytometer. An evaluation study of this low-CD4+ T-lymphocyte blood sample was performed by sending to participating laboratories to investigate the potential use as CD4 EQA material. Our results showed that a magnetic separation technique could be used to prepare low-CD4+ T-lymphocyte blood samples. CD4+ T-lymphocytes in the low-CD4+ T-lymphocyte blood samples ranged from 15 % to 18 % and 160–300 cell/μl, respectively. In addition, CD4 testing of our low-CD4+ T-lymphocyte blood samples could be achieved following both the single- and the dual-platform flow cytometric approaches. A pilot EQA investigation revealed that the CV values of the number and percentage of CD4+ T-lymphocytes were 14 % and 10 %, respectively. Having the low-CD4+ T-lymphocyte blood samples in our CD4 EQA program should ensure reliability and credibility of CD4 testing in Thailand and other resource-poor countries.