Patterns of pseudo-reticulocytosis in malaria: fluorescent analysis with the Cell-Dyn CD4000

This study of Plasmodium falciparum malaria evaluated patterns of fluorescent reticulocyte measurements as determined with the Abbott Cell-Dyn CD4000. The parasitaemia of positive samples (n=180) ranged from 0.04% to 25.5%, with those (19/180) showing gametocytes having lower parasitaemia levels (mean 0.31%, median 0.2%) compared to those that did not (mean 2.59%, median 0.8%). There was a reasonable association (R2=0.60) between parasitaemia level and CD4000 reticulocyte percentages, although there was overall a small statistical bias towards higher parasitaemia estimates determined microscopically. Consistently high immature reticulocyte fraction (IRF) values of >0.5 were observed in cases with a parasitaemia exceeding 5%, while samples with lower parasitaemia levels showed more variable IRF values. Visual examination of CD4000 reticulocyte histograms revealed that 81/100 malaria-positive samples with an IRF above 0.5 showed the presence of a fluorescent population 'spike' consistent with the staining of intracellular malaria parasites. Only three of the 80 malaria-positive samples with an IRF below 0.5, and none of the 237 malaria-negative samples, showed this histogram pattern. These observations indicate that samples with malaria parasites give erroneously high CD4000 reticulocyte estimates that essentially comprise the sum total of true reticulocytes and parasite-infected red cells (pseudo-reticulocytes). This limitation is common to all automated reticulocyte procedures but recognizing the differences between homogenous staining parasitized red cells and heterogeneous staining reticulocytes has potential applications in monitoring parasitaemia levels both at patient presentation and during subsequent treatment.