Impedance-Based Rapid Diagnostic Tool for Single Malaria Parasite Detection.

This paper presents a custom, low-cost electronic system specifically designed for rapid and quantitative detection of the malaria parasite in a blood sample. The system exploits the paramagnetic properties of malaria-infected red blood cells (iRBCs) for their magnetophoretic capture on the surface of a silicon chip. A lattice of nickel magnetic micro-concentrators embedded in a silicon substrate concentrates the iRBCs above coplanar gold microelectrodes separated by 3 µm for their detection through an impedance measurement. The sensor is designed for a differential operation to remove the large contribution given by the blood sample. The electronic readout automatically balances the sensor before each experiment and reaches a resolution of 15 ppm in the impedance measurement at 1 MHz allowing a limit of detection of 40 parasite/µl with a capture time of 10 minutes. For better reliability of the results, four sensors are acquired during the same experiment. We demonstrate that the realized platform can also detect a single infected cell in real experimental conditions, measuring human blood infected by Plasmodium falciparum malaria specie.

Active Opto-Magnetic Biosensing with Silicon Microring Resonators.

Integrated optical biosensors are gaining increasing attention for their exploitation in lab-on-chip platforms. The standard detection method is based on the measurement of the shift of some optical quantity induced by the immobilization of target molecules at the surface of an integrated optical element upon biomolecular recognition. However, this requires the acquisition of said quantity over the whole hybridization process, which can take hours, during which any external perturbation (e.g., temperature and mechanical instability) can seriously affect the measurement and contribute to a sizeable percentage of invalid tests. Here, we present a different assay concept, named Opto-Magnetic biosensing, allowing us to optically measure off-line (i.e., post hybridization) tiny variations of the effective refractive index seen by microring resonators upon immobilization of magnetic nanoparticles labelling target molecules. Bound magnetic nanoparticles are driven in oscillation by an external AC magnetic field and the corresponding modulation of the microring transfer function, due to the effective refractive index dependence on the position of the particles above the ring, is recorded using a lock-in technique. For a model system of DNA biomolecular recognition we reached a lowest detected concentration on the order of 10 pm, and data analysis shows an expected effective refractive index variation limit of detection of 7.5×10-9 RIU, in a measurement time of just a few seconds.

On-chip magnetophoretic capture in a model of malaria-infected red blood cells.

The search for new rapid diagnostic tests for malaria is a priority for developing an efficient strategy to fight this endemic disease, which affects more than 3 billion people worldwide. In this study, we characterize systematically an easy-to-operate lab-on-chip, designed for the magnetophoretic capture of malaria-infected red blood cells (RBCs). The method relies on the positive magnetic susceptibility of infected RBCs with respect to blood plasma. A matrix of nickel posts fabricated in a silicon chip placed face down is aimed at attracting infected cells, while healthy cells sediment on a glass slide under the action of gravity. Using a model of infected RBCs, that is, erythrocytes with methemoglobin, we obtained a capture efficiency of about 70% after 10 min in static conditions. By proper agitation, the capture efficiency reached 85% after just 5 min. Sample preparation requires only a 1:10 volume dilution of whole blood, previously treated with heparin, in a phosphate-buffered solution. Nonspecific attraction of untreated RBCs was not observed in the same time interval.

A Lab-On-chip Tool for Rapid, Quantitative, and Stage-selective Diagnosis of Malaria.

Malaria remains the most important mosquito-borne infectious disease worldwide, with 229 million new cases and 409.000 deaths in 2019. The infection is caused by a protozoan parasite which attacks red blood cells by feeding on hemoglobin and transforming it into hemozoin. Despite the WHO recommendation of prompt malaria diagnosis, the quality of microscopy-based diagnosis is frequently inadequate while rapid diagnostic tests based on antigens are not quantitative and still affected by non-negligible false negative/positive results. PCR-based methods are highly performant but still not widely used in endemic areas. Here, a diagnostic tool (TMek), based on the paramagnetic properties of hemozoin nanocrystals in infected red blood cells (i-RBCs), is reported on. Exploiting the competition between gravity and magnetic forces, i-RBCs in a whole blood specimen are sorted and electrically detected in a microchip. The amplitude and time evolution of the electrical signal allow for the quantification of i-RBCs (in the range 10-105 i-RBC µL-1) and the distinction of the infection stage. A preliminary validation study on 75 patients with clinical suspect of malaria shows on-field operability, without false negative and a few false positive results. These findings indicate the potential of TMek as a quantitative, stage-selective, rapid test for malaria.

On-Chip Selective Capture and Detection of Magnetic Fingerprints of Malaria.

The development of innovative diagnostic tests is fundamental in the route towards malaria eradication. Here, we discuss the sorting capabilities of an innovative test for malaria which allows the quantitative and rapid detection of all malaria species. The physical concept of the test exploits the paramagnetic property of infected erythrocytes and hemozoin crystals, the magnetic fingerprints of malaria common to all species, which allows them to undergo a selective magnetophoretic separation driven by a magnetic field gradient in competition with gravity. Upon separation, corpuscles concentrate at the surface of a silicon microchip where interdigitated electrodes are placed in close proximity to magnetic concentrators. The impedance variation proportional to the amount of attracted particles is then measured. The capability of our test to perform the selective detection of infected erythrocytes and hemozoin crystals has been tested by means of capture experiments on treated bovine red blood cells, mimicking the behavior of malaria-infected ones, and suspensions of synthetic hemozoin crystals. Different configuration angles of the chip with respect to gravity force and different thicknesses of the microfluidic chamber containing the blood sample have been investigated experimentally and by multiphysics simulations. In the paper, we describe the optimum conditions leading to maximum sensitivity and specificity of the test.