This reported the good staging accuracy of PCR (i.e. SE and SP above 80%), but limited utility during post-therapeutic follow-up (specificity and sensitivity varied between 50% and 80%) [57]. These selleck results confirmed previously published data on the strong potential of PCR compared

to other approaches for stage determination, including LATEX/IgM and WBC count, if the presence of parasites in CSF was considered as the only gold standard [113]. However, it is well known that the detection of parasites in CSF is not sensitive enough to be used as a unique staging method. Further studies are needed to evaluate the real added value of PCR compared to the standard stage determination tools. Another aspect that should be taken into account is the low utility of PCR during the post-therapeutic follow-up, since positive results are obtained from patients considered cured [57]. As already highlighted for diagnosis, the application selleck chemical of standard PCR methods in the field is not

practicable. The detection of parasites through amplification of specific DNA sequences in CSF using the LAMP approach is a promising alternative for field application and interesting preliminary results have been reported [114]. An alternative method for easier WBC counts has also been proposed. Based on the observation of the presence of a high number of B-cells expressing CD19 in the CSF of S2 patients, the proposed method consisted Pyruvate dehydrogenase in counting this B-cell population through the formation of rosettes that can be easily visualized and counted by microscopy [115] and [116]. All the biomarkers and tools for the stage determination of HAT described in the previous paragraphs derived from current knowledge of the mechanisms and manifestations of the disease’s progression. However, one of the most common approaches for the discovery of disease biomarkers is the systematic evaluation

of the changes in protein expression between healthy and pathological conditions. As already highlighted, the application of proteomics on sleeping sickness at the host level is an unexplored field. Only one study has been published so far comparing the CSF proteome of S1 and S2 HAT patients [117]. By applying complementary proteomics discovery approaches, it has been shown that a number of host proteins were over-expressed in the CSF of S2 patients. Not surprisingly the 2-DE proteome maps of S2 patients showed a huge increase in the amount of immunoglobulins, in accord with previous knowledge of an elevation of immunoglobulins in CSF with disease progression [85]. Two of the 73 differentially expressed proteins, beta-2-microglobulin and osteopontin, were further confirmed to be accurate discriminators of S1 and S2 patients (AUC% = 91.5 and 84.8, respectively) [117], however after verification other proteins on the list may be found to be more useful.