Abstract
BackgroundWith malaria drug resistance increasing in prevalence and severity, new technologies are needed to aid and improve the accuracy and clinical relevance of laboratory or field testing for malaria drug resistance. This study presents a method based on simple and reagentless spectroscopic measurements coupled with comprehensive spectral interpretation analysis that provides valuable quantitative information on the morphological and compositional responses of Plasmodium falciparum and infected red blood cells (IRBCs) to anti-malarial treatment.MethodsThe changes in the size, internal structure, nucleotide and haemozoin composition of the parasites as well as the morphology (size and shape) and haemoglobin composition of the IRBCs treated with dihydroartemisinin (DHA) and mefloquine (MFQ) were investigated using a spectral interpretation analysis.ResultsDHA treatment reduced the sizes of the parasites and their structural organelles. The haemoglobin composition of the host IRBCs determined from spectroscopic analysis changed negligibly following DHA treatment. MFQ treated parasites grew to the same size as those from parallel non-treated cultures but lacked haemozoin. Lesser deformation of the cell shape and no haemoglobin depletion were detected for the IRBCs of MFQ treated cultures.ConclusionsThe spectroscopic analysis method proved to be sensitive for recognition of the effects of anti-malarial treatment on the structure and composition of the parasites and IRBCs. The method can have significant potential for research and clinical applications such as evaluating patient specimens for drug action, drug effects or for therapeutic monitoring.
Highlights
With malaria drug resistance increasing in prevalence and severity, new technologies are needed to aid and improve the accuracy and clinical relevance of laboratory or field testing for malaria drug resistance
Techniques based on polymerase chain reaction (PCR) offer superior sensitivity, ability to detect mixed species infection and the ability to differentiate among four species of Plasmodium but they require considerable cost of equipment as well as storage and maintenance requirements for reagents [2,4]
Quantitative PCR has been repeatedly demonstrated to be more accurate than microscopy for the detection of malaria at very low levels of parasitaemia [7,8] and it disagrees with microscopy for parasitaemia counts at higher parasitaemia levels [8]
Summary
With malaria drug resistance increasing in prevalence and severity, new technologies are needed to aid and improve the accuracy and clinical relevance of laboratory or field testing for malaria drug resistance. Microscopy has long been the “gold standard” as a tool for the research based characterization of malaria infected red blood cells (IRBCs) and for the detection of malaria parasites in the field [1,2,3,4,5] Because this technique is constrained by substantial technical and economic hurdles such as operator training, time needed to perform a test, and labor intensiveness, there is an ongoing search for alternatives. Quantitative PCR has been repeatedly demonstrated to be more accurate than microscopy for the detection of malaria at very low levels of parasitaemia [7,8] and it disagrees with microscopy for parasitaemia counts at higher parasitaemia levels [8] It is not straightforward and requires complicated analysis to assess the sensitivity of the malaria parasites to anti-malarials using immunochromatography or molecular methods [9,10,11] and is time-consuming when using classical in vivo studies [12]
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