RESUMO
A novel method for the in vitro detection of the protozoan Plasmodium, the causative agent of malaria, has been developed. It comprises a protocol for cleanup of whole blood samples, followed by direct ultraviolet laser desorption (LD) time-of-flight mass spectrometry. Intense ion signals are observed from intact ferriprotoporphyrin IX (heme), sequestered by malaria parasites during their growth in human red blood cells. The LD mass spectrum of the heme is structure-specific, and the signal intensities are correlated with the sample parasitemia (number of parasites per unit volume of blood). Parasitemia levels on the order of 10 parasites/microL blood can be unambiguously detected by this method. Consideration of laser beam parameters (spot size, rastering across the sample surface) and actual sample consumption suggests that the detection limits can be further improved by at least an order of magnitude. The influence of experimental factors, such as desorbed ion polarity, laser exposure and fluence, sample size, and parasite growth stage, on the threshold for parasite detection is also addressed.