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Micromagnetic resonance relaxometry for rapid label-free malaria diagnosis

Abstract

We report a new technique for sensitive, quantitative and rapid detection of Plasmodium spp.–infected red blood cells (RBCs) by means of magnetic resonance relaxometry (MRR). During the intraerythrocytic cycle, malaria parasites metabolize large amounts of cellular hemoglobin and convert it into hemozoin crystallites. We exploit the relatively large paramagnetic susceptibility of these hemozoin particles, which induce substantial changes in the transverse relaxation rate of proton nuclear magnetic resonance of RBCs, to infer the 'parasite load' in blood. Using an inexpensive benchtop 0.5-Tesla MRR system, we show that with minimal sample preparatory steps and without any chemical or immunolabeling, a parasitemia level of fewer than ten parasites per microliter in a volume below 10 μl of whole blood is detected in a few minutes. We demonstrate this method both for cultured Plasmodium falciparum parasites and in vivo with Plasmodium berghei–infected mice.

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Figure 1: Home-built, low-cost MRR system, experimental setup and the conceptual idea of magnetic susceptibility of infected RBCs.
Figure 2: P.falciparum infection and the R2 responses as measured by the MRR system.
Figure 3: In vivo P.berghei infection and the R2 responses as measured by the MRR system.
Figure 4: Performance comparison for MRR and microscopy technique in early stage of P. berghei ANKA infection detection.

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Acknowledgements

This work is supported by the National Research Foundation Singapore under its SMART Centre, BioSystems and Micromechanics IRG and Infectious Disease IRG. W.K.P. acknowledges support from the SMART Postdoctoral Research Fellows Programme and SMART Ignition Grant (ING 11025-BIO(IGN)) and K.R. Roy for culturing and preparing the sample parasites. T.F.K. and C.S.N. acknowledge financial support from the Singapore-MIT Alliance (SMA) Graduate Fellowship.

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Contributions

W.K.P. and J.H. conceived the original idea and designed the study. W.K.P. built the whole experimental setup, designed the protocol and performed MRR measurements of P. falciparum and P. berghei. L.C. assisted in radiofrequency probe design. C.S.N., W.K.P., A.A.S.B. and L.C. were involved in preparing P. falciparum parasite samples. T.F.K., C.S.N., W.K.P. and Y.H. worked on the in vivo mice studies (day-to-day measurements). C.S.N., W.K.P., L.C. and T.F.K. contributed equally to the blinded mouse studies. P.R.P., N.-T.N. and J.H. supervised the mouse work. W.K.P., P.R.P., J.H., T.F.K. and C.S.N. wrote the paper, and all the authors checked through the manuscript and analyzed the data.

Corresponding authors

Correspondence to Weng Kung Peng or Peter Rainer Preiser or Jongyoon Han.

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Competing interests

W.K.P. and J.H. have filed patents with the World Intellectual Property Office on the technique discussed here: a technology patent describing the strategies for miniaturization of the devices and an application patent describing its usage as a malaria screening tool (ref. 9).

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Supplementary Tables 1–5, Supplementary Figures 1–10 Supplementary Results and Supplementary Discussion (PDF 3343 kb)

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Peng, W., Kong, T., Ng, C. et al. Micromagnetic resonance relaxometry for rapid label-free malaria diagnosis. Nat Med 20, 1069–1073 (2014). https://doi.org/10.1038/nm.3622

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